Therapy for inhibition of single-stranded rna virus replication

ABSTRACT

Pharmaceutical compositions showing the ability to inhibit or suppress replication of a filovirus in an individual are disclosed. The disclosed compositions are useful for treating, reventing, or reducing the spread of infections by filovirus. A method includes administering at least one agent of the present disclosure to an individual infected with or exposed to a filovirus, wherein the step of administering is carried out for a suitable time period so that the individual is treated; and determining whether the individual has been treated, wherein the step of determining includes one of measuring an inhibition in viral replication, measuring a decrease in viral load, or reducing at least one symptom associated with the filovirus.

RELATED APPLICATIONS

This application claims the benefit of and priority to U.S. ProvisionalPatent Application No. 62/068,465, filed Oct. 24, 2014, U.S. ProvisionalPatent Application No. 62/068,469, filed Oct. 24, 2014, U.S. ProvisionalPatent Application No. 62/068,477, filed Oct. 24, 2014, U.S. ProvisionalPatent Application No. 62/068,487, filed Oct. 24, 2014, U.S. ProvisionalPatent Application No. 62/068,492, filed Oct. 24, 2014, and U.S.Provisional Patent Application No. 62/188,030, filed Jul. 2, 2015, theentirety of these applications is hereby incorporated herein byreference.

BACKGROUND

RNA viruses can be classified according to the sense or polarity oftheir RNA into negative-sense (−) and positive-sense (+) RNA viruses.The largest family of viruses is the single stranded negative-sense (−)RNA (“ssRNA”) family of viruses. Their viral RNA genome cannot bedirectly translated, instead the (−) strand is complementary to theviral mRNAs that need to be produced and translated into viral proteins.At the time of this disclosure, one order and eight families arerecognized in this group. There are also a number of unassigned speciesand genera:

Order Mononegavirales

-   -   Family Bornaviridae—Borna disease virus    -   Family Filoviridae—includes Ebola virus, Marburg virus    -   Family Paramyxoviridae—includes Measles virus, Mumps virus,        Nipah virus, Hendra virus, RespiratorySyncytial Virus (RSV), and        Newcastle Disease Virus (NDV)    -   Family Rhabdoviridae—includes Rabies virus    -   Family Nyamiviridae—includes Nyavirus

Unassigned families:

-   -   Family Arenaviridae—includes Lassa virus    -   Family Bunyaviridae—includes Hantavirus, Crimean-Congo        hemorrhagic fever    -   Family Ophioviridae    -   Family Orthomyxoviridae—includes Influenza viruses

Unassigned genera:

-   -   Genus Deltavirus—includes Hepatitis D virus    -   Genus Dichorhavirus    -   Genus Emaravirus    -   Genus Nyavirus—includes Nyamanini and Midway viruses    -   Genus Tenuivirus    -   Genus Varicosavirus

Unassigned species:

-   -   Taastrup virus    -   Sclerotinia sclerotiorum negative-stranded RNA virus 1

Despite decades of efforts by researchers to develop an effective,approved, and available filovirus treatment for individuals, currentlythere are no United States Food and Drug Administration-approvedvaccines or therapeutics for treatment of infection with filovirusdiseases.

SUMMARY

Therapies for the inhibition of single-stranded RNA virus replicationare disclosed herein. Compositions and methods for treating symptomaticand/or asymptomatic infections of negative-sense single-stranded RNAviruses including, but not limited to, Bornaviridae, Filoviridae,Paramyxoviridae, Rhabdoviridae, Nyamiviridae or any combination thereof,are disclosed herein. In an embodiment, therapy for inhibition of anEbola virus is disclosed herein. In an embodiment, therapy forinhibition of Marburg virus is disclosed herein.

A method of the present invention includes administering a compound toan individual infected with or exposed to a filovirus, wherein the stepof administering is carried out for a suitable time period so that theindividual is treated, and wherein the compound is represented byformula I:

-   -   or a pharmaceutically acceptable salt thereof, wherein:    -   R₁ is phenyl, 4-chlorophenyl or 4-fluorophenyl,    -   R₂ is 4-pyridyl, optionally substituted with up to 4 groups that        are independently (C₁-C₆) alkyl, halogen, haloalkyl,        —OC(O)(C₁-C₆ alkyl), —C(O)O(C₁-C₆ alkyl), —CONR′R″, —OC(O)NR′R″,        —NR′C(O)R″, —CF₃, —OCF₃, —OH, C₁-C₆ alkoxy, hydroxyalkyl, —CN,        —CO₂H, —SH, —S-alkyl, —SOR′R″, —SO₂R′, —NO₂, or NR′R″, wherein        R′ and R″ are independently H or (C₁-C₆) alkyl, and wherein each        alkyl portion of a substituent is optionally further substituted        with 1, 2, or 3 groups independently selected from halogen, CN,        OH, and NH₂,    -   R₄ is H or alkyl, and    -   n is 1 or 2; and

determining whether the individual has been treated, wherein the step ofdetermining comprising one of measuring an inhibition in viralreplication, measuring a decrease in viral load, or reducing at leastone symptom associated with the filovirus. In an embodiment, thecompound of formula I is:

In an embodiment, the compound of formula I is administered at a dailydosage ranging from about 2.5 mg/kg to about 22.5 mg/kg. In anembodiment, the compound of formula I is administered at a daily dosageranging from about 3.5 mg/kg to about 21.5 mg/kg. In an embodiment, thecompound of formula I is administered at a daily dosage ranging fromabout 4.5 mg/kg to about 20.5 mg/kg. In an embodiment, the compound offormula I is administered at a daily dosage ranging from about 5.5 mg/kgto about 19.5 mg/kg. In an embodiment, the compound of formula I isadministered at a daily dosage ranging from about 6.5 mg/kg to about18.5 mg/kg. In an embodiment, the compound of formula I is administeredat a daily dosage ranging from about 7.5 mg/kg to about 17.5 mg/kg. Inan embodiment, the compound of formula I is administered at a dailydosage ranging from about 8.5 mg/kg to about 16.5 mg/kg. In anembodiment, the compound of formula I is administered at a daily dosageranging from about 9.5 mg/kg to about 15.5 mg/kg. In an embodiment, thecompound of formula I is administered at a daily dosage ranging fromabout 10.5 mg/kg to about 14.5 mg/kg. In an embodiment, the compound offormula I is administered at a daily dosage ranging from about 11.5mg/kg to about 13.5 mg/kg. In an embodiment, the determining stepincludes measuring, at at least two different times during the suitabletime period, the viral load using a nucleic acid amplification basedtest. In an embodiment, the inhibition in viral replication or thedecrease in viral load is at least 10% as determined using a nucleicacid amplification based test. In an embodiment, the individual is ahuman. In an embodiment, the filovirus is Ebola virus or Marburg virus.In an embodiment, the filovirus is Ebola virus. In an embodiment, thecompound of formula I is present as a solid dosage form. In anembodiment, the solid dosage form is a capsule. In an embodiment, themethod further includes administering at least one antibiotic to theindividual infected with or exposed to the filovirus for the suitabletime period, wherein the combination of the at least one antibiotic andthe compound of formula I produce a synergistic effect. In anembodiment, the at least one antibiotic is selected from one ofclarithromycin or rifabutin.

A method of the present invention includes administering at least twoantibiotics to an individual infected with or exposed to a filovirus,wherein the step of administering is carried out for a suitable timeperiod so that the individual is treated; and determining whether theindividual has been treated, wherein the step of determining includesone of measuring an inhibition in viral replication, measuring adecrease in viral load, or reducing at least one symptom associated withthe filovirus. In an embodiment, at least one of the antibiotics is amacrolide antibiotic. In an embodiment, at least one of the antibioticsis a rifamycin antibiotic. In an embodiment, the antibiotics areclarithromycin and rifabutin. In an embodiment, clarithromycin isadministered at a daily dosage ranging from about 2.5 mg/kg to about21.5 mg/kg. In an embodiment, clarithromycin is administered at a dailydosage ranging from about 3.5 mg/kg to about 20.5 mg/kg. In anembodiment, clarithromycin is administered at a daily dosage rangingfrom about 4.5 mg/kg to about 19.5 mg/kg. In an embodiment,clarithromycin is administered at a daily dosage ranging from about 5.5mg/kg to about 18.5 mg/kg. In an embodiment, clarithromycin isadministered at a daily dosage ranging from about 6.5 mg/kg to about17.5 mg/kg. In an embodiment, rifabutin is administered at a dailydosage ranging from about 7.5 mg/kg to about 16.5 mg/kg. In anembodiment, clarithromycin is administered at a daily dosage rangingfrom about 8.5 mg/kg to about 15.5 mg/kg. In an embodiment,clarithromycin is administered at a daily dosage ranging from about 9.5mg/kg to about 14.5 mg/kg. In an embodiment, rifabutin is administeredat a daily dosage ranging from about 10.5 mg/kg to about 13.5 mg/kg. Inan embodiment, rifabutin is administered at a daily dosage ranging fromabout 0.5 mg/kg to about 7.5 mg/kg. In an embodiment, rifabutin isadministered at a daily dosage ranging from about 1.5 mg/kg to about 6.5mg/kg. In an embodiment, rifabutin is administered at a daily dosageranging from about 2.5 mg/kg to about 5.5 mg/kg. In an embodiment, thedetermining step includes measuring, at at least two different timesduring the suitable time period, the viral load using a nucleic acidamplification based test. In an embodiment, the inhibition in viralreplication or the decrease in viral load is at least 10% as determinedusing a suitable assay. In an embodiment, the individual is a human. Inan embodiment, the filovirus is Ebola virus or Marburg virus. In anembodiment, the filovirus is Ebola virus.

DETAILED DESCRIPTION

As used herein, the term “agent” refers to a compound having apharmacological activity—an effect of the agent on an individual. Theterms “agent,” “compound,” and “drug” are used interchangeably herein.

A “patient” or an “individual” refers to any animal, such as a primate.In an embodiment, the primate is a non-human primate. In an embodiment,the primate is a human primate. Any animal can be treated using themethods and composition of the present invention.

As used herein, the term “synergistic effect” refers to the coordinatedor correlated action of two or more agents of the present invention sothat the combined action is greater than the sum of each actingseparately. In an embodiment, agents of the present invention, whenadministered together as part of a treatment regimen, provide atherapeutic synergy without accompanying synergistic side effects (e.g.,but not limited to, cross-reacting agents).

As used herein, the term “treat” is meant to administer one or moreagents of the present invention to measurably inhibit the replication ofa virus in vitro or in vivo, to measurably decrease the load of a virusin a cell in vitro or in vivo, or to reduce at least one symptomassociated with having a filovirus-mediated disease in a patient.Desirably, the inhibition in replication or the decrease in viral loadis at least 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, 95%, 97%, 98%, 99%, as determined using asuitable assay. Assays that monitor replication of viruses include, butare not limited to, cytopathic viral assays, reporter-virus andreporter-cell assays, viral replicon assays, and gene-targeted viralassays. In an embodiment, an assay that measures CD8 T cell-mediatedinhibition of filovirus replication is used to measure the slow or stopin the replication of a virus. Viral load testing can be carried outusing nucleic acid amplification based tests (NATs or NAATs) andnon-nucleic acid-based tests on blood plasma samples to determine thequantity of virus in a given volume including viral RNA levels in plasmaand tissue and total viral DNA. Alternatively, in certain embodiments,treatment is observed by a trained physician as an appreciable orsubstantial relief of symptoms in a patient with a filovirus-mediateddisease. Typically, a decrease in viral replication is accomplished byreducing the rate of RNA polymerization, RNA translation, proteinprocessing or modification, or by reducing the activity of a moleculeinvolved in any step of viral replication (e.g., proteins or coded bythe genome of the virus or host important for viral replication). In anembodiment, the term “treat” refers to the ability of an agent or agentsof the present invention to inhibit or suppress replication of a virus,such as an RNA virus. In an embodiment, the term “treat” refers to theability of an agent or agents of the present invention to inhibit thecytopathic effect during a RNA virus infection.

By an “effective amount” is meant the amount of an agent or agents ofthe present invention, alone or in combination with another therapeuticregimen, required to treat a patient with a viral disease (e.g., anyvirus described herein including an Ebola virus or Marburg virus) in aclinically relevant manner. A sufficient effective amount of an agent oragents used to practice the present invention for therapeutic treatmentof conditions caused by a virus varies depending upon the manner ofadministration, the age, body weight, and general health of the patient.Ultimately, the prescribers will decide the appropriate amount anddosage regimen. In a combination therapy of the invention, the effectiveamount of an agent may be less than the effective amount if the agentwere administered in a non-combinatorial (single-agent) therapy.Additionally, an effective amount may be an amount of an agent in acombination therapy of the invention that is safe and efficacious in thetreatment of a patient having a viral disease over each agent alone asdetermined and approved by a regulatory authority (such as the U.S. Foodand Drug Administration).

By “more effective” is meant that a treatment exhibits greater efficacy,or is less toxic, safer, more convenient, or less expensive than anothertreatment with which it is being compared. Efficacy may be measured by askilled practitioner using any standard method that is appropriate for agiven indication.

By a “filovirus” is meant a virus belonging to the family Fillovirdae.Exemplary filoviruses are Ebola virus and Marburg virus.

“Ebola” or “Ebola hemorrhagic fever” is a disease caused by infectionwith one of the Ebola virus strains. Ebola can cause disease in humansand nonhuman primates (monkeys, gorillas, and chimpanzees). Eboladisease in humans is caused by four of five viruses in the genusEbolavirus. The four are Bundibugyo virus (BDBV), Sudan virus (SUDV),Tai Forest virus (TAFV), and one called, simply, Ebola virus (EBOV,formerly Zaire Ebola virus). The fifth virus, Reston virus (RESTV), isnot thought to cause disease in humans, but has caused disease in otherprimates. These five viruses are closely related to marburgviruses.Marburg virus disease (MVD) is a severe illness of humans and non-humanprimates caused by either of the two marburgviruses, Marburg virus andRavn virus.

As used herein, the term “a suitable period of time” refers to theperiod of time starting when a patient begins treatment for a diagnosisof ssRNA viral infection (e.g., but not limited to, Ebola) using amethod of the present disclosure, throughout the treatment, and up untilwhen the patient stops treatment due to either a reduction in symptomsassociated with ssRNA viral infection (e.g., but not limited to, Ebola)or due to a laboratory diagnosis indicating that the ssRNA viralinfection (e.g., but not limited to, Ebola) is under control. In anembodiment, a suitable period of time is one (1) week. In an embodiment,a suitable period of time is between one (1) week and two (2) weeks. Inan embodiment, a suitable period of time is two (2) weeks. In anembodiment, a suitable period of time is between two (2) weeks and three(3) weeks. In an embodiment, a suitable period of time is three (3)weeks. In an embodiment, a suitable period of time is between three (3)weeks and four (4) weeks. In an embodiment, a suitable period of time isfour (4) weeks. In an embodiment, a suitable period of time is betweenfour (4) weeks and five (5) weeks. In an embodiment, a suitable periodof time is five (5) weeks. In an embodiment, a suitable period of timeis between five (5) weeks and six (6) weeks. In an embodiment, asuitable period of time is six (6) weeks. In an embodiment, a suitableperiod of time is between six (6) weeks and seven (7) weeks. In anembodiment, a suitable period of time is seven (7) weeks. In anembodiment, a suitable period of time is between seven (7) weeks andeight (8) weeks. In an embodiment, a suitable period of time is eight(8) weeks.

As used herein, the term “cytopathic effects” refers to the changes incell morphology due to a viral infection.

As used herein, the terms “cytopathogenesis” or “pathogenesis” includesinhibition of host cell gene expression and includes other cellularchanges that contribute to viral pathogenesis in addition to thosechanges that are visible at the microscopic level.

The term “in vitro” as used herein refers to procedures performed in anartificial environment, such as for example, without limitation, in atest tube or cell culture system. The skilled artisan will understandthat, for example, an isolate SK enzyme may be contacted with amodulator in an in vitro environment. Alternatively, an isolated cellmay be contacted with a modulator in an in vitro environment.

The term “in vivo” as used herein refers to procedures performed withina living organism such as, without limitation, a human, monkey, mouse,rat, rabbit, bovine, equine, porcine, canine, feline, or primate.

Some small viruses carry their genome as single-stranded DNA (ssDNA)molecules. These viruses have a simple genome: one gene for a viralnucleocapsid protein and another gene for a DNA replication enzyme. Thevirus with a ssDNA genome also faces a serious replication problem inthe host cell. When introduced into cells, these genomes cannot be usedto make viral proteins because the only template for transcription isdouble-stranded DNA. For this reason, the first step after infection isthe conversion of the viral ssDNA into dsDNA using host cell DNApolymerase. In some of these viruses, the 3′ end of the viral DNA foldsback and forms dsDNA by base-pairing with an internal sequence. In thisway, the primer is built into the genome and the 3′ end can be extendedto create dsDNA that serves as a template for transcription. Theresulting transcripts are translated to make the viral proteins, thereplicated viral DNA is converted back into a ssDNA genome, and thevirion is packaged for export.

RNA viruses can be classified according to the sense or polarity oftheir RNA into negative-sense (−) and positive-sense (+), or ambisenseRNA viruses. Positive-sense viral RNA is similar to mRNA and thus can beimmediately translated by the host cell. Negative-sense viral RNA iscomplementary to mRNA and thus must be converted to positive-sense RNAby an RNA polymerase before translation. As such, purified RNA of apositive-sense virus can directly cause infection though it may be lessinfectious than the whole virus particle. Purified RNA of anegative-sense virus is not infectious by itself as it needs to betranscribed into positive-sense RNA; each virion can be transcribed toseveral positive-sense RNAs. Ambisense RNA viruses resemblenegative-sense RNA viruses, except they also translate genes from thepositive strand. Examples of positive-strand RNA viruses include, butare not limited to, polio virus, Coxsackie virus, and echovirus.Examples of negative-strand RNA viruses include, but are not limited to,influenza virus, measles viruses, and rabies virus.

The largest family of viruses is the (−) ssRNA family of viruses. Theirviral RNA genome cannot be directly translated, instead the (−) strandis complementary to the viral mRNAs that need to be produced andtranslated into viral proteins. Nature has created hundreds of different(−) ssRNA viruses ranging from the measles and influenza viruses to therabies and Ebola viruses. Members of this class of virus include Ebolavirus and members of the influenza family of viruses.

Ebola, previously known as Ebola hemorrhagic fever, is a disease causedby infection with one of the Ebola virus strains. Ebola can causedisease in humans and nonhuman primates (monkeys, gorillas, andchimpanzees). Ebola disease in humans is caused by four of five virusesin the genus Ebolavirus. The four are Bundibugyo virus (BDBV), Sudanvirus (SUDV), Tai Forest virus (TAFV), and one called, simply, Ebolavirus (EBOV, formerly Zaire Ebola virus). The fifth virus, Reston virus(RESTV), is not thought to cause disease in humans, but has causeddisease in other primates. These five viruses are closely related tomarburgviruses. Currently, no specific therapy is available that hasdemonstrated efficacy in the treatment of Ebola.

Ebolaviruses contain single-strand, non-infectious RNA genomes.Ebolavirus genomes are approximately 19 kilobase pairs long and containseven genes in the order 3′-UTR—NP-VP35-VP40-GP-VP30-VP24-L-5′-UTR. Thegenomes of the five different ebolaviruses (BDBV, EBOV, RESTV, SUDV, andTAFV) differ in sequence and the number and location of gene overlaps.In general, ebolavirions are 80 nanometers (nm) in width and may be aslong as 14,000 nm. In general, the median particle length ofebolaviruses ranges from 974 to 1,086 nm (in contrast to marburgvirions,whose median particle length was measured at 795-828 nm), but particlesas long as 14,000 nm have been detected in tissue culture.

The viral matrix protein 40 (VP40) is the most abundant protein found inthe virions, in infected cells, and also inside the viral nucleocapsid.The nucleoprotein (NP) is associated with the viral genome and assembledinto a helical nucleocapside (NC) along with polymerase cofactor (VP35),the transcription activator (VP30), and the RNA-dependent RNA polymerase(L). The viral proteins that comprise the NC catalyze the replicationand transcription of the viral genome. A minor viral matrix protein,VP24 is also required for NC assembly. If NP is expressed alone incells, it assembles together with cellular RNA to form a loose coil-likestructure. When NP is co-expressed with VP24 and VP35, NC-likestructures are formed in the cytoplasm that are morphologicallyindistinguishable from those seen in infected cells. It has been shownthat VP24 and the viral matrix protein VP40 reduce the transcription andreplication efficiencies of the EBOV genome, suggesting that VP24 andVP40 are important for the conversion from a transcription andreplication-competent NC to one that is ready for viral assembly. VP40plays a role in the formation and release of the enveloped, filamentousvirus-like particles (VLPs) even when expressed alone. NC-likestructures are incorporated into VLPs when VP40 is co-expressed with NP,VP35, and VP24, suggesting that a direct interaction between VP40 and NPis important for the recruitment of NC-like structures to the buddingsite, the plasma membrane. The interaction between VP40 and NP is alsorequired for the formation of condensed NC-like structures. GP is asurface glycoprotein that forms spikes on virions and plays a crucialrole in virus entry into cells by mediating receptor binding and fusion.

The ebolavirus life cycle begins with virion attachment to specificcell-surface receptors, followed by fusion of the virion envelope withcellular membranes and the concomitant release of the virus nucleocapsidinto the cytosol. Ebolavirus' structural glycoprotein (known as GP1,2)is responsible for the virus' ability to bind to and infect targetedcells. The viral RNA polymerase, encoded by the L gene, partiallyuncoats the nucleocapsid and transcribes the genes into positive-strandmRNAs, which are then translated into structural and nonstructuralproteins. The most abundant protein produced is the nucleoprotein, whoseconcentration in the cell determines when L switches from genetranscription to genome replication. Replication results in full-length,positive-strand antigenomes that are, in turn, transcribed intonegative-strand virus progeny genome copy. Newly synthesized structuralproteins and genomes self-assemble and accumulate near the inside of thecell membrane. Virions bud off from the cell, gaining their envelopesfrom the cellular membrane they bud from. The mature progeny particlesthen infect other cells to repeat the cycle. The Ebola virus geneticsare difficult to study due to its virulent nature.

Ebola is a filamentous, enveloped, negative-sense RNA strand virus inthe family of Filoviridae. The RNA-dependent RNA-polymerase of Ebolavirus shares significant sequence homology to other negative-strand RNAviruses, required for both viral transcription and replication of theviral genome. However, RNA-dependent RNA-polymerase requires a hostfactor and viral proteins cooperating to accomplish replication andtranscription. One of the reasons why Ebola is so deadly is due to itsability to circumvent the immune system while at the same timepro-actively destroying the human body, as a result the immune system isnot able to gather a cohesive effort to fight off the disease. Duringthe infection, monocytes/macrophages in the lymphoid tissues are earlyand sustained targets of this deadly virus. During the viral infection,large amounts of proinflammatory cytokines such as tumor necrosis factor(TNF-α) are secreted from infected macrophages and cause disruption ofthe endothelial barrier. Macrophages and Dendritic cells play a centralrole in inducing the observed clinical feature of Ebola's hemorrhagicfever. Secreted cytokines, chemokines, and other mediators alter theblood vessel functions, promote and recruit an influx of inflammatorycells, including additional monocytes/macrophages to the site of theinfection. The virus released from the infected macrophages anddendritic cells spread to similar cells throughout the body and toparenchymal cells in many organs, resulting in multifocal tissuenecrosis. The ability of the host to develop an effective adaptiveimmune response is weakened by massive lymphocyte apoptosis, aphenomenon also seen in bacterial sepsis. Ebola infection also induceslymphocyte apoptosis, although the virus does not replicate inlymphocytes. Recent studies indicate that NK (Natural Killer) cells andCD4+ and CD8+ lymphocytes are the principal cell types affected inEbola-infected macaques monkeys.

An Ebola subject who shows symptoms (i.e., is symptomatic) including,but not limited to, high fever, headache, joint and muscle aches, sorethroat, weakness, stomach pain, lethargy, and lack of appetite, canundergo blood and/or tissue tests to confirm an Ebola diagnosis. AnEbola subject who does not show symptoms (i.e., is asymptomatic) canundergo blood and/or tissue tests to confirm an Ebola diagnosis. Theseasymptomatic subjects may have markers in their blood indicating theycarry the disease, but they are totally asymptomatic.

A ssRNA virus infected subject who shows symptoms (i.e., is symptomatic)including, but not limited to, high fever, headache, joint and muscleaches, sore throat, weakness, stomach pain, lethargy, and lack ofappetite, can undergo blood and/or tissue tests to confirm a diagnosisof a ssRNA virus infection. A ssRNA virus infected subject who does notshow symptoms (i.e., is asymptomatic) can undergo blood and/or tissuetests to confirm a diagnosis of ssRNA virus infection. Theseasymptomatic subjects may have markers in their blood indicating theycarry the disease, but they are totally asymptomatic.

In an embodiment, a subject can be tested for a ssRNA viral infection(e.g., but not limited to, Ebola) within a few days after symptomsbegin, or after treatment according to the present disclosure, bycollecting a blood or other body fluid sample and testing the sample fordetection of viral antigens or RNA in blood and other body fluids using,for example, an antigen-capture enzyme-linked immunosorbent assay(ELISA), using an IgM ELISA (to determine whether the subject has IgMantibodies), using an IgG ELISA (to determine whether the subject hasIgG antibodies), using polymerase chain reaction (PCR), or by virusisolation.

The present disclosure identifies agents and combinations of agentshaving inhibitory activity against a model filovirus. The presentinvention features compositions and methods for the treatment offilovirus-mediated disease, e.g., one caused by an Ebola virus orMarburg virus.

In an embodiment, the present disclosure describes a method for treatinga patient with a filovirus-mediated disease, for example a diseasecaused by Ebola virus or Marburg virus. The method includesadministering to the patient a first agent selected from the agents ofTable 1, or an analog thereof, in an amount that is effective to treatthe patient. In an embodiment, the method further includes administeringa second agent selected from the agents of Table 1. In an embodiment,the method further includes administering a third agent selected fromthe agents of Table 1.

TABLE 1 Clofazamine Rifabutin Clarithromycin Brivudine AryladamantaneUpamostat Compounds

When the methods include administering to a patient more than one activeagent, the agents may be administered within 7, 6, 5, 4, 3, 2 or 1 days;within 24, 12, 6, 5, 4, 3, 2 or 1 hours, within 60, 50, 40, 30, 20, 10,5 or 1 minutes; or substantially simultaneously. The methods of theinvention may include administering one or more agents to the patient byoral, systemic, parenteral, topical, intravenous, inhalational, orintramuscular administration. In an embodiment, the methods of theinvention include administering one or more agents to the patient byoral administration.

In an embodiment, the present disclosure describes a compositionincluding two or more agents selected from the agents of Table 1. In anembodiment, the two or more agents are present in amounts that, whenadministered together to a patient with a filovirus-mediated diseasesuch as a disease caused by Ebola virus or Marburg virus, are effectiveto treat the patient. In an embodiment, the composition consists ofactive ingredients and excipients, and the active ingredients consist oftwo or more agents selected from agents of Table 1.

Active ingredients or agents useful in the invention include thosedescribed herein in any of their pharmaceutically acceptable forms,including isomers, salts, solvates, and polymorphs thereof, as well asracemic mixtures and prodrugs.

According to aspects illustrated herein, a treatment regimen of thepresent disclosure is suitable to inhibit the viral replicationmachinery of a ssRNA virus, and is also suitable to inhibit thecytopathic effect during a ssRNA virus infection.

Small Hsps interact with a large number of client proteins that areessential to many cellular processes. For example, Hsp90 interacts withover 200 polypeptides in order to modulate their activity and/or halflife. Hsp90, HspB1, and probably other small Hsps, are global regulatorsof cell systems. Hsp90 is a host factor for the replication of negativestrand viruses and is responsible for proteins folding properly,intracellular disposition, stabilizing proteins against heat stress, andalso proteolytic turnover of many essential regulators of cell growthand differentiation.

Brivudine (bromovinyldeoxyuridine or BVDU for short) interacts with twophenylalanine residues (Phe29 and Phe33) in the N-terminal domain ofHspB1. The drug's full chemical description is(E)-5-(2-bromovinyl)-2-deoxyuridine. Brivudine is a nucleoside analoguetargeting two viral enzymes: deoxythymidine kinases and polymerases.Brivudine is able to be incorporated into viral DNA, and then blocks theaction of DNA polymerases, thus inhibiting viral replication. An oralformulation of the present disclosure including brivudine can be used anapproach for a therapeutic ssRNA viral infection. The active compound isthe 5′-triphosphate of BVDU, which is formed in subsequentphosphorylations by viral thymidine kinase and presumably by nucleosidediphosphate kinase. Brivudine can bind in vitro to the heat shockprotein HSPB1 and inhibits interaction with its binding partners.Brivudine has properties against HSP27, HSP70 and HSP90.

A non-limiting example of the prodrug form of BVDU is represented byFormula I:

Brivudine is represented by Formula 2:

A solid oral dosage composition of the present disclosure includes(E)-5-(2-bromovinyl)2′-deoxyuridine (BVDU), a salt thereof, or BVDU inprotected or in prodrug form, and at least one conventional carrier andmay include at least one auxiliary material. BVDU may be present in anamount effective to produce a concentration of 0.02 μg/ml to 10.0 μg/mlin blood.

According to aspects illustrated herein, there is disclosed acomposition comprising brivudine (BVDU), an active metabolite of BVDU, asalt thereof, or BVDU in protected or in prodrug form wherein brivudine(BVDU), an active metabolite of BVDU, a salt thereof, or BVDU inprotected or in prodrug form is present in an amount that, whenadministered to a patient with a filovirus-mediated disease, iseffective to treat the patient. In an embodiment, the filovirus is Ebolavirus or Marburg virus.

According to aspects illustrated herein, a method for treating a patienthaving filovirus-mediated disease includes administering to the patienta composition comprising brivudine (BVDU), an active metabolite of BVDU,a salt thereof, or BVDU in protected or in prodrug form in an amounteffective to treat the patient. In an embodiment, the filovirus is Ebolavirus or Marburg virus.

Upamostat (“WX-671” or “Mesupron”) inhibits the urokinase-typeplasminogen activator (uPA) system. Upamostat is a serine proteaseinhibitor. After oral administration, serine protease inhibitor WX-671is converted to the activeNα-(2,4,6-triisopropylphenylsulfonyl)-3-amidino-(L)-phenylalanine-4-ethoxycarbonylpiperazide(“WX-UK1”), which inhibits several serine proteases, particularly uPA.The serine protease inhibitor upamostat can potentially inhibitreplication of viral RNA. An oral formulation of the present disclosureincluding upamostat can be used an approach for a therapeutic ssRNAviral infection. The drug's full chemical description is (S)-ethyl4-(3-(3-(N-hydroxycarbamimidoyl)phenyl)-2-(2,4,6-triisopropylphenylsulfonamido)propanoyl)piperazine-1-carboxylate. In an embodiment of the present disclosure,upamostat is administered orally at a dose of about 0.5-to about 1.1mg/kg. In an embodiment, upamostat is administered orally at a dailydose of between about 200 mg to 400 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 150 mg to 550 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 200 mg to 550 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 250 mg to 550 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 300 mg to 550 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 350 mg to 550 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 400 mg to 550 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 450 mg to 550 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 500 mg to 550 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 200 mg to 550 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 200 mg to 500 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 200 mg to 450 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 200 mg to 350 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 200 mg to 300 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 200 mg to 250 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 500 mg to 1000 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 750 mg to 1000 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 500 mg to 750 mg.

Upamostat is represented by Formula 3:

In an embodiment of the present disclosure, upamostat is administeredorally at a dose of about 0.5-to about 1.1 mg/kg. In an embodiment,upamostat is administered orally at a daily dose of between about 200 mgto 400 mg. In an embodiment, upamostat is administered orally at a dailydose of between about 150 mg to 550 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 200 mg to 550 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 250 mg to 550 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 300 mg to 550 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 350 mg to 550 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 400 mg to 550 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 450 mg to 550 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 500 mg to 550 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 200 mg to 550 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 200 mg to 500 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 200 mg to 450 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 200 mg to 350 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 200 mg to 300 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 200 mg to 250 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 500 mg to 1000 mg. In an embodiment, upamostat isadministered orally at a daily dose of between about 750 mg to 1000 mg.In an embodiment, upamostat is administered orally at a daily dose ofbetween about 500 mg to 750 mg.

Clofazimine is a riminophenazine compound which can accumulate to veryhigh concentrations in tissues. Clofazimine can inhibit bacterial growthby inducing apoptosis of host cells. Treatment with clofazimine mayresult in highly condensed chromatin within the nucleus, indicatingmacrophages undergoing apoptosis. Macrophages are responsible for aseries of biochemical products with potent immunomodulatory activities.Additionally, clofazimine may stimulate the activity of variousreticuloendothelial phagocytic cells. The cells, primarily monocytes andmacrophages, accumulate in lymph nodes and the spleen, in connectivetissues and in the liver, in lungs, and the central nervous system(microglia). The mononuclear phagocyte system is an integral part ofboth humoral and cell-mediated immunity and has an important roleagainst microorganisms, including mycobacteria, fungi, and viruses.

According to aspects illustrated herein, there is disclosed acomposition comprising clofazamine, wherein clofazamine is present in anamount that, when administered to a patient with a filovirus-mediateddisease, is effective to treat the patient. In an embodiment, thefilovirus is Ebola virus or Marburg virus. According to aspectsillustrated herein, a method for treating a patient havingfilovirus-mediated disease includes administering to the patient acomposition comprising clofazamine in an amount effective to treat thepatient. In an embodiment, the filovirus is Ebola virus or Marburgvirus.

In an embodiment, the dosage of clofazimine per day is from about 15mg/day to about 120 mg/day till recovery. In an embodiment, the maximumdosage of clofazimine is 80-120 mg/day till recovery. In an embodiment,clofazimine is administered orally as a 50 mg tablet twice per day. Inan embodiment, clofazimine is administered orally as a 100 mg tabletonce per day. In an embodiment, clofazimine is administered as acomponent of a solid oral dosage form comprising from 10 mg to 16 mg ofclofazimine per dosage form, for up to six times per day. In anembodiment, a method for treating a ssRNA viral infection, such asEbola, comprises concomitantly administering: i) up to 120 mg/day of anoral clofazimine dosage form, ii) up to 450 mg/day of an oral rifabutindosage form, iii) up to 1000 mg/day clarithromycin as either anintravenous infusion or a solid oral dosage form, and iv) up to 600mg/day of an oral dosage form comprising brivudine, an active metaboliteof brivudine, a salt thereof, or BVDU in protected or in prodrug form,wherein the administration is for a period of time until the subject isfree and clear of a ssRNA viral infection, such as Ebola. Clofazimine isrepresented by Formula 4:

A ssRNA viral infection, such as Ebola virus, induces massive lymphocyteapoptosis, which is thought to prevent the development of a functionaladaptive immune response. Rifabutin may be effective in inhibiting thecytopathic effect of the ssRNA viral infection, such as Ebola viruswhich destroys the T lymphocytes. Rifabutin interferes with RNAreplication machinery by inhibiting RNA synthesis (transcription) and,consequently, the initiation of a new round of DNA replication.

According to aspects illustrated herein, there is disclosed acomposition comprising rifabutin, wherein rifabutin is present in anamount that, when administered to a patient with a filovirus-mediateddisease, is effective to treat the patient. In an embodiment, thefilovirus is Ebola virus or Marburg virus. According to aspectsillustrated herein, a method for treating a patient havingfilovirus-mediated disease includes administering to the patient acomposition comprising rifabutin in an amount effective to treat thepatient. In an embodiment, the filovirus is Ebola virus or Marburgvirus.

In an embodiment, the dosage of rifabutin per day is from about 80mg/day to about 480 mg/day. In an embodiment, the maximum dosage ofrifabutin is 480 mg/day till recovery. In an embodiment, rifabutin isadministered orally as a 150 mg tablet twice per day. In an embodiment,rifabutin is administered orally as a 300 mg tablet once per day. In anembodiment, rifabutin is administered as a component of a solid oraldosage form comprising from 45 mg to 60 mg of rifabutin per dosage form,for up to six times per day. In an embodiment, a method for treating assRNA viral infection, such as Ebola virus, comprises concomitantlyadministering: i) up to 450 mg/day of an oral rifabutin dosage form, ii)up to 120 mg/day of an oral clofazimine dosage form, iii) up to 1000mg/day clarithromycin as either an intravenous infusion or a solid oraldosage form, and iv) up to 600 mg/day of an oral dosage form comprisingbrivudine, an active metabolite of brivudine, a salt thereof, or BVDU inprotected or in prodrug form. Rifabutin is represented by Formula 5:

According to aspects illustrated herein, there is disclosed acomposition comprising clarithromycin, wherein clarithromycin is presentin an amount that, when administered to a patient with afilovirus-mediated disease, is effective to treat the patient. In anembodiment, the filovirus is Ebola virus or Marburg virus. According toaspects illustrated herein, a method for treating a patient havingfilovirus-mediated disease includes administering to the patient acomposition comprising clarithromycin in an amount effective to treatthe patient. In an embodiment, the filovirus is Ebola virus or Marburgvirus.

In an embodiment, the dosage of clarithromycin per day is from about 180mg/day to about 1000 mg/day till recovery. In an embodiment, the maximumdosage of clarithromycin is 980-1000 mg/day till recovery. In anembodiment, two doses of 500 mg clarithromycin is administered as anintravenously (IV) infusion, using a solution concentration of about 2mg/ml. 1 gram daily of clarithromycin can be administered as anintravenously (IV) infusion for a period of from two days to five days.In an embodiment, 1 gram daily of clarithromycin can be administered asan intravenously (IV) infusion for a period of three days. In anembodiment, clarithromycin is administered orally as a 500 mg tablettwice per day. In an embodiment, clarithromycin is administered as acomponent of a solid oral dosage form comprising from 95 mg to 125 mg ofclarithromycin per dosage form. The solid dosage form can beadministered up to twelve times per day. In an embodiment, a method fortreating a ssRNA viral infection, such as Ebola virus, comprisesintravenously administering for a first period of time, such as forexample, from 2 days to 5 days, 1 gram daily of clarithromycin followedby orally administering for a second period of time, such as from theend of the first period of time until the subject is free and clear of assRNA viral infection, such as Ebola virus, 1 gram daily ofclarithromycin. In an embodiment, a method for treating a ssRNA viralinfection, such as Ebola virus, comprises concomitantly administering:i) up to 1 gram/day of clarithromycin, ii) up to 600 mg/day of an oraldosage form comprising brivudine, an active metabolite of brivudine, asalt thereof, or BVDU in protected or in prodrug form, iii) up to 120mg/day of an oral clofazimine dosage form, and iv) up to 450 mg/day ofan oral rifabutin dosage form, wherein the administration is for aperiod of time until the subject is free and clear of a ssRNA viralinfection, such as Ebola virus. Clarithromycin is represented by Formula6:

Proinflammatory cytokines such as TNF-α, IL-1β, IL-6, IL-8, IL-12 andIFN-γ, increase vascular permeability, promote vascular leakage and therecruitment of neutrophils at the inflammatory site and stimulate theproduction of acute-phase proteins. In a clinical state, they inducefever or hypothermia, and peripheral shock. Co-administration ofclarithromycin with clofazimine and/or rifabutin in animal models iseffective at reducing pro-inflammatory cytokines such as TNF-α and IL-6specifically. During viral infection large amounts of proinflammatorycytokines like TNF-α are secreted from macrophages. The combination ofthe above drugs could effectively inhibit proinflammatory cytokines anddecrease the permeability of the vascular endothelium, which facilitatesthe virus' entry into endothelial cells.

Clarithromycin is an effective immunomodulator. Rifabutin enhances theantimicrobial and intercellular effects of clarithromycin. Additionally,clarithromycin as an immunomodulator can have considerable efficacy forpatients with sepsis, as sometimes present in a ssRNA viral infection,such as Ebola virus infection. In an embodiment, clarithromycin isadministered orally to a subject as a component of a solid oral dosageform. In an embodiment, clarithromycin is administered as an intravenousinfusion to a subject. In an embodiment, clarithromycin may beadministered as an intravenous infusion to a subject in conjunction withan oral dosage form of Brivudine (BVDU), an active metabolite of BVDU, asalt thereof, or BVDU in protected or in prodrug form. In an embodiment,clarithromycin may be administered as an intravenous infusion to asubject in conjunction with an oral dosage form of upamostat.

According to aspects illustrated herein, there is disclosed acomposition comprising rifabutin and clarithromycin, wherein rifabutinand clarithromycin are present in amounts that, when administeredtogether to a patient with a filovirus-mediated disease, are effectiveto treat the patient. In an embodiment, the filovirus is Ebola virus orMarburg virus. According to aspects illustrated herein, a method fortreating a patient having filovirus-mediated disease includesadministering to the patient a composition comprising rifabutin andclarithromycin in amounts effective to treat the patient. In anembodiment, the filovirus is Ebola virus or Marburg virus.

According to aspects illustrated herein, there is disclosed acomposition comprising clofazamine, rifabutin and clarithromycin,wherein clofazamine, rifabutin and clarithromycin are present in amountsthat, when administered together to a patient with a filovirus-mediateddisease, are effective to treat the patient. In an embodiment, thefilovirus is Ebola virus or Marburg virus. According to aspectsillustrated herein, a method for treating a patient havingfilovirus-mediated disease includes administering to the patient acomposition comprising clofazamine, rifabutin and clarithromycin inamounts effective to treat the patient. In an embodiment, the filovirusis Ebola virus or Marburg virus.

In an embodiment, rifabutin, clarithromycin, and clofazimine areadministered as a single solid oral dosage form. In an embodiment, asolid oral dosage form of the present disclosure comprises rifabutin,clarithromycin, clofazimine, and a pharmaceutically acceptable carrier,wherein the amount of clofazimine is 5-18% w/w relative to the amount ofclarithromycin (such as, 7-16%, 9-14%, 9-12%, 10-15%, or 0-11% w/w) and10-25% w/w relative to the amount of rifabutin (such as, 12-25%, 12-23%,15-25%, 15-23%, 18-25%, 18-23%, 20-25%, 20-23%, or 21-23%).

In an embodiment, a solid oral dosage form of the present disclosurecomprises rifabutin, clarithromycin, and clofazimine in an8-10:18-20:1-2.5 w/w/w ratio (for example, a 8.5-9.5:18.5-19.5:1.5-2.5w/w/w ratio or a 9:19:2 ratio, wherein each variable is free to vary±0.5 or 0.25). In an embodiment, a solid oral dosage form of the presentdisclosure comprises rifabutin, clarithromycin, and clofazimine in abouta 9:19:2 w/w/w ratio, wherein each of the variables are free to vary ±2,1, 0.5, or 0.25 (e.g., 9±0.5:19±5:21.0.5). For example in an embodiment,a solid oral dosage form of the present disclosure comprises 90 mgrifabutin (±30, 20, 10, 5, 2, or 1 mg), 190 mg clarithromycin (±60, 40,20, 10, 5, 2, or 1 mg), and 20 mg clofazimine (±10, 7, 5, 2, or 1 mg).In an embodiment, a solid oral dosage form of the present disclosurecomprises 45 mg rifabutin (±15, 10, 7, 5, 2, or 1 mg), 95 mgclarithromycin (±30, 20, 10, 5, 2, or 1 mg), and 10 mg clofazimine (±6,5, 2, or 1 mg).

In embodiment, a solid oral dosage form of the present disclosurefurther comprises an absorption enhancer that may improvebioavailability of one or more of the active ingredients. The amount ofabsorption enhancer may between 300-700% w/w relative to the amount ofclofazimine including 400-600% or 450-550% or 475-525%. In certainembodiments, the absorption enhancer is polyethylene glycol (PEG), forexample, polyethylene glycol having an average molecular weight ofbetween 200-20,000 including between 1000-15000 or 5000-12000 or7000-9000 or 7500-8500, for example PEG 8000).

In embodiment, a solid oral dosage form of the present disclosurefurther comprises one or more additional excipients, such asMCC-Tabulose type 200, Mg Stearate, SLS-Emal 10Pwd HD, a polysorbate(such as, polysorbate 80), or a combination thereof, including all ofthese. In some instances, the present compositions include bothpolyethylene glycol and a polysorbate, such as polysorbate 80, whereinthe amount of polysorbate is 30-120% w/w relative to the amount ofclofazimine (such as 50-100%, 50-85%, or 60-75%).

In an embodiment, a solid oral dosage form of the present disclosurefurther comprises one or more additional excipients, such asMicrocrystalline cellulose (MCC) TABULOSE® SC 200), Mg Stearate, SodiumLauryl Sulfate (SLS) EMAL® 10Pwd HD, a polysorbate (such as, polysorbate80), or a combination thereof, including all of these. In someinstances, the present compositions include both polyethylene glycol anda polysorbate, such as polysorbate 80, wherein the amount of polysorbateis 30-120% w/w relative to the amount of clofazimine (such as 50-100%,50-85%, or 60-75%).

In an embodiment, a solid oral dosage form of the present disclosure isavailable in the form of a tablet or a capsule containing an active in apowdered form. In an embodiment, a solid oral dosage form of the presentdisclosure is in the form of a tablet or a capsule containing an activein a microencapsulated form. In an embodiment, a solid oral dosage formof the present disclosure is in the form of a tablet or a capsulecontaining an active in a microgranulated form.

In an embodiment, a solid oral dosage form of the present disclosure isavailable in the form of a tablet comprising at least one of rifabutin,clarithromycin, and clofazimine in a powdered form. In some instancestwo or all of rifabutin, clarithromycin, and clofazimine are in apowdered form. In an embodiment, a solid oral dosage form of the presentdisclosure is in the form of a tablet or a capsule comprising at leastone of rifabutin, clarithromycin, and clofazimine in a microencapsulatedform. In some instances, two or all of rifabutin, clarithromycin, andclofazimine are in a microencapsulated form. In an embodiment, a solidoral dosage form of the present disclosure is in the form of a tablet ora capsule comprising at least one of rifabutin, clarithromycin, andclofazimine in a powdered form, and the remaining agents present in amicroencapsulated form. In an embodiment, a solid oral dosage form ofthe present disclosure is in the form of a tablet or a capsulecomprising one or more of rifabutin, clarithromycin, and clofazimine ina microgranulated form. In an embodiment, a solid oral dosage form ofthe present disclosure is in the form of a tablet comprising one or moreof rifabutin, clarithromycin, and clofazimine within a capsule, acapsule containing one or more of rifabutin, clarithromycin, andclofazimine within a tablet, a capsule containing one or more ofrifabutin, clarithromycin, and clofazimine within an outer capsulecontaining the other agents, or any combination of the above. In anembodiment, a solid oral dosage form of the present disclosure comprisesan inner capsule containing rifabutin, within an outer capsulecontaining clarithromycin and clofazimine, wherein clarithromycin andclofazimine may be present in powdered, microencapsulated, ormicrogranulated forms.

Surprisingly, concomitant administration of multiple drugs disclosedherein, including, but not limited to concomitant administration ofbrivudine with at least one of clofazimine, rifabutin, clarithromycin,or upamostat, can inhibit the replication machinery of a ssRNA virusinfection so as to hinder, restrain or prevent viral infection.

Surprisingly, concomitant administration of multiple drugs disclosedherein, including, but not limited to concomitant administration ofbrivudine with at least one of clofazimine, rifabutin, clarithromycin,or upamostat, can inhibit the replication machinery of a ssRNA virusinfection so as to hinder, restrain or prevent viral infection.

Surprisingly, concomitant administration of multiple drugs disclosedherein, including, but not limited to concomitant administration ofbrivudine with at least one of clofazimine, rifabutin, clarithromycin,or upamostat, can inhibit the cytopathic effect of a ssRNA virusinfection so as to hinder, restrain or prevent degenerative changes orabnormalities.

According to aspects illustrated herein, there is disclosed acomposition comprising rifabutin, clarithromycin, and brivudine whereinrifabutin, clarithromycin and brivudine are present in amounts that,when administered together to a patient with a filovirus-mediateddisease, are effective to treat the patient. In an embodiment, thefilovirus is Ebola virus or Marburg virus.

According to aspects illustrated herein, a method for treating a patienthaving filovirus-mediated disease includes administering to the patienta composition comprising rifabutin, clarithromycin and Brivudine inamounts effective to treat the patient. In an embodiment, the filovirusis Ebola virus or Marburg virus.

According to aspects illustrated herein, a method for treating a subjecthaving a ssRNA viral infection includes concomitant administration of atherapeutically effective amount of brivudine (BVDU), an activemetabolite of BVDU, a salt thereof, or BVDU in protected or in prodrugform; a therapeutically effective amount of rifabutin; and atherapeutically effective amount of clarithromycin.

According to aspects illustrated herein, a method for treating a subjecthaving a ssRNA viral infection includes concomitant administration asfollows: orally administering a tablet comprising a therapeuticallyeffective amount of brivudine (BVDU), an active metabolite of BVDU, asalt thereof, or BVDU in protected or in prodrug form; orallyadministering a capsule or tablet comprising rifabutin; and orallyadministering a capsule or tablet comprising clarithromycin, wherein therifabutin and the clarithromycin may be present in the same capsule ortablet.

According to aspects illustrated herein, a method for treating a subjecthaving a ssRNA viral infection includes concomitant administration asfollows: orally administering a therapeutically effective amount ofbrivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDUin protected or in prodrug form for a suitable period of time; andadministering a therapeutically effective amount of at least one ofclofazimine, rifabutin or clarithromycin, for a suitable period of time.In an embodiment, a therapeutically effective amount of BVDU is up to600 mg/day for an adult. In an embodiment, the 600 mg is administeredonce daily as a single oral dosage form. In an embodiment, the 600 mg isadministered as a 150 mg single oral dosage form taken four times daily.In an embodiment, a therapeutically effective amount of BVDU is up to500 mg/day for an adult. In an embodiment, the 500 mg is administeredonce daily as a single oral dosage form. In an embodiment, the 500 mg isadministered as a 125 mg single oral dosage form taken four times daily.In an embodiment, the therapeutically effective amount of clofazimine isfrom about 50 mg to about 300 mg daily for an adult. In an embodiment,clofazimine is administered orally as a solid dosage form one or moretimes per day. In an embodiment, the therapeutically effective amount ofrifabutin is from about 45 mg to about 480 mg daily for an adult. In anembodiment, rifabutin is administered orally as a solid dosage form oneor more times per day. In an embodiment, the therapeutically effectiveamount of clofazimine is from about 50 mg to about 300 mg daily for anadult. In an embodiment, clofazimine is administered orally as a soliddosage form one or more times per day.

According to aspects illustrated herein, a method for treating a subjecthaving a ssRNA viral infection includes concomitant administration asfollows: orally administering a therapeutically effective amount ofbrivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDUin protected or in prodrug form for a suitable period of time; andorally administering a therapeutically effective amount of a solid oraldosage form that comprises at least one of clofazimine or rifabutin fora suitable period of time; and administering a therapeutically effectiveamount of clarithromycin as an intravenous infusion for a suitableperiod of time. In an embodiment, a therapeutically effective amount ofBVDU is up to 600 mg/day for an adult. In an embodiment, the 600 mg isadministered once daily as a single oral dosage form. In an embodiment,the 600 mg is administered as a 150 mg single oral dosage form takenfour times daily. In an embodiment, a therapeutically effective amountof BVDU is up to 500 mg/day for an adult. In an embodiment, the 500 mgis administered once daily as a single oral dosage form. In anembodiment, the 500 mg is administered as a 125 mg single oral dosageform taken four times daily. In an embodiment, a therapeuticallyeffective amount of clarithromycin is up to 1 gram daily for an adult.In an embodiment, two doses of 500 mg clarithromycin is administered asan IV infusion, using a solution concentration of about 2 mg/ml. 1 gramdaily of clarithromycin can be administered as an IV infusion for aperiod of from two days to five days. In an embodiment, 1 gram daily ofclarithromycin can be administered as an IV infusion for a period ofthree days. In an embodiment, a therapeutically effective amount ofrifabutin is up to 480 mg daily for an adult. In an embodiment,rifabutin is administered orally as a tablet one or more times per day.In an embodiment, rifabutin is administered as a component of a solidoral dosage form comprising from 45 mg to 60 mg of rifabutin per dosageform. A solid dosage form comprising from 45 mg to 60 mg of rifabutincan be administered up to twelve times per day. In an embodiment, atherapeutically effective amount of clofazimine is from about 50 mg toabout 300 mg daily for an adult. In an embodiment, clofazimine isadministered orally as a solid dosage form one or more times per day. Inan embodiment, clofazimine is administered as a component of a solidoral dosage form comprising from 10 mg to 16 mg of clofazimine perdosage form.

According to aspects illustrated herein, a method for treating a subjecthaving a ssRNA viral infection includes concomitant administration asfollows: orally administering a therapeutically effective amount ofbrivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDUin protected or in prodrug form for a suitable period of time; andadministering a therapeutically effective amount of clarithromycin as anintravenous infusion for a suitable period of time. In an embodiment, atherapeutically effective amount of BVDU is up to 600 mg/day for anadult. In an embodiment, the 600 mg is administered once daily as asingle oral dosage form. In an embodiment, the 600 mg is administered asa 150 mg single oral dosage form taken four times daily. In anembodiment, a therapeutically effective amount of BVDU is up to 500mg/day for an adult. In an embodiment, the 500 mg is administered oncedaily as a single oral dosage form. In an embodiment, the 500 mg isadministered as a 125 mg single oral dosage form taken four times daily.In an embodiment, the therapeutically effective amount of clarithromycinis up to 1 gram daily for an adult. The clarithromycin is administeredas an intravenous (IV) infusion. In an embodiment, two doses of 500 mgclarithromycin is administered as an IV infusion, using a solutionconcentration of about 2 mg/ml. 1 gram daily of clarithromycin can beadministered as an IV infusion for a period of from two days to fivedays. In an embodiment, 1 gram daily of clarithromycin can beadministered as an IV infusion for a period of three days.

According to aspects illustrated herein, a method for treating a subjecthaving a ssRNA viral infection includes concomitant administration asfollows: orally administering a therapeutically effective amount ofbrivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDUin protected or in prodrug form for a suitable period of time; orallyadministering a therapeutically effective amount of a solid oral dosageform that comprises clarithromycin for a suitable period of time; orallyadministering a therapeutically effective amount of a solid oral dosageform that comprises rifabutin for a suitable period of time; and orallyadministering a therapeutically effective amount of a solid oral dosageform that comprises clofazimine, for a suitable period of time. In anembodiment, a therapeutically effective amount of BVDU is up to 600mg/day for an adult. In an embodiment, the 600 mg is administered oncedaily as a single oral dosage form. In an embodiment, the 600 mg isadministered as a 150 mg single oral dosage form taken four times daily.In an embodiment, a therapeutically effective amount of BVDU is up to500 mg/day for an adult. In an embodiment, the 500 mg is administeredonce daily as a single oral dosage form. In an embodiment, the 500 mg isadministered as a 125 mg single oral dosage form taken four times daily.In an embodiment, a therapeutically effective amount of clarithromycinis up to 1 gram daily for an adult. In an embodiment, clarithromycin isadministered orally as a 500 mg tablet twice per day. In an embodiment,a therapeutically effective amount of rifabutin is up to 480 mg dailyfor an adult. In an embodiment, rifabutin is administered orally as atablet one or more times per day. In an embodiment, rifabutin isadministered as a component of a solid oral dosage form comprising from45 mg to 60 mg of rifabutin per dosage form. A solid dosage formcomprising from 45 mg to 60 mg of rifabutin can be administered up totwelve times per day. In an embodiment, a therapeutically effectiveamount of clofazimine is from about 50 mg to about 300 mg daily for anadult. In an embodiment, clofazimine is administered orally as a soliddosage form one or more times per day. In an embodiment, clofazimine isadministered as a component of a solid oral dosage form comprising from10 mg to 16 mg of clofazimine per dosage form.

In an embodiment, clofazimine and brivudine are administered orally as asolid dosage form one or more times per day. In an embodiment,clofazimine, rifabutin, and brivudine are administered orally as a soliddosage form one or more times per day. In an embodiment, rifabutin andbrivudine are administered orally as a solid dosage form one or moretimes per day. In an embodiment, clarithromycin, rifabutin, andbrivudine are administered orally as a solid dosage form one or moretimes per day. In an embodiment, clarithromycin, clofazimine, andbrivudine are administered orally as a solid dosage form one or moretimes per day. In an embodiment, upamostat, clofazimine, and brivudineare administered orally as a solid dosage form one or more times perday. In an embodiment, upamostat, rifabutin, clofazimine, and brivudineare administered orally as a solid dosage form one or more times perday. In an embodiment, upamostat, rifabutin, and brivudine areadministered orally as a solid dosage form one or more times per day. Inan embodiment, upamostat, rifabutin, clarithromycin, and brivudine areadministered orally as a solid dosage form one or more times per day. Inan embodiment, upamostat, brivudine, clarithromycin, and clofazimine areadministered orally as a solid dosage form one or more times per day. Inan embodiment, upamostat, brivudine, clarithromycin, rifabutin, andclofazimine are administered orally as a solid dosage form one or moretimes per day.

In an embodiment, the rifabutin, clarithromycin and clofazimine areadministered as a single solid oral dosage form. In some instances, therifabutin, clarithromycin, and clofazimine are co-administered once eachday for a first period of treatment (for example, 1-3 weeks, including 1week, 2 weeks or three weeks) in the following amounts: (i) 80-100 mgrifabutin (such as, 85-95 mg or 90 mg±1.5 mg), (ii) 180-200 mgclarithromycin (such as, 185-195 mg or 190 mg±2 mg), and (iii) 15-25 mgclofazimine (such as 17-23 mg or 20±1 mg). The method may furtherinclude the step of linearly increasing the amounts of the rifabutin,clarithromycin, and clofazimine while maintaining a 8-10:18-20:1-2.5w/w/w ratio (for example, a 8.5-9.5:18.5-19.5:1.5-2.5 w/w/w ratio or a9:19:2 ratio, wherein each variable is free to vary ±0.5 or 0.25 ratio)for a second period of treatment (for example, from 4-10 weeks). In anembodiment, the linearly increasing amounts of the rifabutin,clarithromycin, and clofazimine do not exceed maximum amounts of (i)420-480 mg rifabutin (such as, 440-460 mg or 450 mg), 920-980 mgclarithromycin (such as, 940-960 mg or 950 mg), and (iii) 80-120 mgclofazimine (such as, 90-110 mg or 100 mg) during the second period oftreatment. In certain instances, the linearly increasing amounts ofrifabutin, clarithromycin, and clofazimine comprise: a) (i) 160-200 mgrifabutin (such as, 170-190 mg or 180 mg±2 mg), (ii) 360-400 mgclarithromycin (such as, 370-390 mg or 380 mg±2 mg), and (iii) 30-50 mgclofazimine (such as, 35-45 mg or 40 mg±1 mg) once each day for twoweeks; b) (i) 250-290 mg rifabutin (such as, 260-280 mg or 270 mg±2 mg),(ii) 550-590 mg clarithromycin (such as, 560-580 mg or 570±2 mg), and(iii) 50-70 mg clofazimine (such as, 55-65 mg or 60 mg±1.5 mg) once eachday for two weeks; c) (i) 340-380 mg rifabutin (such as, 350-370 mg or360 mg±2 mg), (ii) 740-780 mg clarithromycin (such as 750-770 mg or 760mg±2 mg), and (iii) 60-100 mg clofazimine (such as, 70-90 mg or 80mg±1.5 mg) once each day for two weeks; and d) (i) 420-480 mg rifabutin(such as, 440-460 mg or 450 mg±2 mg), (ii) 920-980 mg clarithromycin(such as, 940-960 mg or 950 mg±2 mg), and (iii) 80-120 mg clofazimine(such as, 90-110 mg or 100 mg±1.5 mg) once each day for a week. Incertain embodiments, the method further includes, following step d)above, the step of simultaneously co-administering (i) 420-480 mgrifabutin (such as, 440-460 mg or 450 mg±2 mg), (ii) 920-980 mgclarithromycin (such as, 940-960 mg or 950 mg±2 mg), and (iii) 80-120 mgclofazimine (such as, 90-110 mg or 100 mg±1.5 mg) once each day for athird period of treatment. In some embodiments, the third period oftreatment is 1, 2, 4, 6, 8, 12 weeks; 3, 6, or 12 months or longer. Inone embodiment the third period of treatment continues until either areduction in symptoms associated with a ssRNA viral infection or due toa laboratory diagnosis indicating that the a ssRNA viral disease isunder control.

In an embodiment, the single solid oral dosage form includes rifabutin;clarithromycin; clofazimine; an absorption enhancer; and apharmaceutically acceptable carrier, wherein the pharmaceuticalcomposition is a solid oral dosage form, wherein the absorption enhanceris between 300% and 700% w/w relative to the amount of clofazimine, andwherein an amount of clofazimine is 10-15% w/w relative to an amount ofclarithromycin and 20-25% w/w relative to an amount of rifabutin.

In an embodiment, the single solid oral dosage form includes rifabutin;clarithromycin; clofazimine; polyethylene glycol; and a pharmaceuticallyacceptable carrier, wherein the pharmaceutical composition is a solidoral dosage form, wherein the polyethylene glycol, (i) has an averagemolecular weight of between 1000-15000 Daltons, and (ii) is between 300%and 700% w/w relative to the amount of clofazimine, and wherein anamount of clofazimine is 10-15% w/w relative to an amount ofclarithromycin and 20-25% w/w relative to an amount of rifabutin.

Small heat shock proteins (small Hsps) are stress-induced molecularchaperones that act as holdases towards polypeptides that have losttheir folding in stress conditions or consequently of mutations in theircoding sequence. A cellular protection against the deleterious effectsmediated by damaged proteins is thus provided to cells. These chaperonesare also highly expressed in response to protein conformational andinflammatory diseases and cancer pathologies. Through specific andreversible modifications in their phospho-oligomeric organization, smallHsps can chaperone appropriate client proteins in order to provide cellswith resistance to different types of injuries or pathologicalconditions. By helping cells to better cope with their pathologicalstatus, their expression can be either beneficial, such as in diseasescharacterized by pathological cell degeneration, or deleterious whenthey are required for tumor cell survival. Moreover, small Hsps areactively released by cells and can act as immunogenic molecules thathave dual effects depending on the pathology. Five families of Hsps areinduced by stress: the 70 kDa (HspA-Hsp70) family, the 20-30 kDa(HspB-small Hsps, sHsps) family, the 90 kDa (HspC-Hsp90) family, the 60kDa (HspD-Hsp60) family, and the HspH (large Hsps) family.

An aryladamantane compound of the present invention has been shown to becapable of selectively inhibiting SK2 activity in vitro. Without beingbound by theory, it is believed that inhibition of sphingosine kinase(SK) may impair viral protein expression and infectious virus productionfrom cells expressing a cellular protein that acts as a receptor forEbola virus and Marburg virus. According to aspects illustrated herein,there is disclosed a composition comprising an aryladamantane compound,wherein the aryladamantane compound is present in an amount that, whenadministered to a patient with a filovirus-mediated disease, areeffective to treat the patient. In an embodiment, the filovirus is Ebolavirus or Marburg virus. According to aspects illustrated herein, amethod for treating a patient having filovirus-mediated disease includesadministering to the patient a composition comprising an aryladamantanecompound in an amount effective to treat the patient. In an embodiment,the filovirus is Ebola virus or Marburg virus.

The symbol “—” in general represents a bond between two atoms in thechain. Thus CH₃—O—CH₂—CH(R_(i))—CH₃ represents a2-substituted-1-methoxypropane compound. In addition, the symbol “—”represents the point of attachment of the substituent to a compound.Thus for example aryl(C₁-C₆)alkyl- indicates an alkylaryl group, such asbenzyl, attached to the compound at the alkyl moiety.

Where multiple substituents are indicated as being attached to astructure, it is to be understood that the substituents can be the sameor different. Thus for example “R_(m) optionally substituted with 1, 2or 3 R_(q) groups” indicates that R_(m) is substituted with 1, 2, or 3R_(q) groups where the R_(q) groups can be the same or different.

The phrase “optionally substituted” is used interchangeably with thephrase “substituted or unsubstituted”. Unless otherwise indicated, anoptionally substituted group may have a substituent at eachsubstitutable position of the group, and each substituent is independentof the other.

As used herein, the terms “halogen” or “halo” indicate fluorine,chlorine, bromine, or iodine.

The term “heteroatom” means nitrogen, oxygen or sulfur and includes anyoxidized form of nitrogen and sulfur, and the quaternized form of anybasic nitrogen. Also the term “nitrogen” includes a substitutablenitrogen in a heterocyclic ring. As an example, in a saturated orpartially unsaturated ring having 0-3 heteroatoms selected fromnitrogen, oxygen or sulfur, the nitrogen may be N (as in3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR⁺ (as inN-substituted pyrrolidinyl).

The term “alkyl”, as used herein alone or as part of a larger moiety,refers to a saturated aliphatic hydrocarbon including straight chain,branched chain or cyclic (also called “cycloalkyl”) groups. Examples ofalkyl groups include methyl, ethyl, propyl, isopropyl, butyl, iso-, sec-and tert-butyl, pentyl, hexyl, heptyl, 3-ethylbutyl, and the like.Preferably, the alkyl group has 1 to 20 carbon atoms (whenever anumerical range, e.g. “1-20”, is stated herein, it means that the group,in this case the alkyl group, may contain 1 carbon atom, 2 carbon atoms,3 carbon atoms, etc. up to and including 20 carbon atoms). Morepreferably, it is a medium size alkyl having 1 to 10 carbon atoms. Mostpreferably, it is a lower alkyl having 1 to 4 carbon atoms. Thecycloalkyl can be monocyclic, or a polycyclic fused system. Examples ofcycloalkyl groups include cyclopropyl, cyclobutyl, cycolpentyl,cyclohexyl, cycloheptyl, cyclooctyl, and adamantyl. The alkyl orcycloalkyl group may be unsubstituted or substituted with 1, 2, 3 ormore substituents. Examples of such substituents including, withoutlimitation, halo, hydroxy, amino, alkoxy, alkylamino, dialkylamino,cycloalkly, aryl, aryloxy, arylalkyloxy, heterocyclic radical, and(heterocyclic radical)oxy. Examples include fluoromethyl, hydroxyethyl,2,3-dihydroxyethyl, (2- or 3-furanyl)methyl, cyclopropylmethyl,benzyloxyethyl, (3-pyridinyl)methyl, (2-thienyl)ethyl, hyroxypropyl,aminocyclohexyl, 2-dimethylaminobutyl, methoxymethyl, N-pyridinyl ethyl,and diethylaminoethyl.

The term “cycloalkylalkyl”, as used herein alone or as part of a largermoiety, refers to a C₃-C₁₀ cycloalkyl group attached to the parentmolecular moiety through an alkyl group, as defined above. Examples ofcycloalkylalkyl groups include cyclopropylmethyl and cyclopentylethyl,

The term “alkenyl”, as used herein alone or as part of a larger moiety,refers to an aliphatic hydrocarbon having at least one carbon-carbondouble bond, including straight chain, branched chain or cyclic groupshaving at least one carbon-carbon double bond. Preferably, the alkenylgroup has 2 to 20 carbon atoms. More preferably, it is a medium sizealkenyl having 2 to 10 carbon atoms. Most preferably, it is a loweralkenyl having 2 to 6 carbon atoms. The alkenyl group may beunsubstituted or substituted with 1, 2, 3 or more substituents. Examplesof such substituents including, without limitation halo, hydroxy, amino,alkoxy, alkylamino, dialkylamino, cycloalkly, aryl, aryloxy,arylalkyloxy, heterocyclic radical, and (heterocyclic radical)oxy.Depending on the placement of the double bond and substituents, if any,the geometry of the double bond may be entgegen (E) or zusammen (Z),cis, or trans. Examples of alkenyl groups include ethenyl, propenyl,cis-2-butenyl, trans-2-butenyl, and 2-hyroxy-2-propenyl.

The term “alkynyl”, as used herein alone or as part of a larger moiety,refers to an aliphatic hydrocarbon having at least one carbon-carbontriple bond, including straight chain, branched chain or cyclic groupshaving at least one carbon-carbon triple bond. Preferably, the alkynylgroup has 2 to 20 carbon atoms. More preferably, it is a medium sizealkynyl having 2 to 10 carbon atoms. Most preferably, it is a loweralkynyl having 2 to 6 carbon atoms. The alkynyl group may beunsubstituted or substituted with 1, 2, 3 or more substituents. Examplesof such substituents including, without limitation, halo, hydroxy,amino, alkoxy, alkylamino, dialkylamino, cycloalkly, aryl, aryloxy,arylalkyloxy, heterocyclic radical, and (heterocyclic radical)oxy.Examples of alkynyl groups include ethynyl, propynyl, 2-butynyl, and2-hyroxy-3-butylnyl.

The term “alkoxy”, as used herein alone or as part of a larger moiety,represents an alkyl group of indicated number of carbon atoms attachedto the parent molecular moiety through an oxygen bridge. Examples ofalkoxy groups include, for example, methoxy, ethoxy, propoxy andisopropoxy. Alkoxy radicals may be further substituted with one or morehalo atoms, such as fluoro, chloro or bromo, to provide “haloalkoxy”radicals. Examples of such radicals include fluoromethoxy,chloromethoxy, trifluoromethoxy, and fluoroethoxy.

The term “aryl”, as used herein alone or as part of a larger moiety,refers to an aromatic hydrocarbon ring system containing at least onearomatic ring. The aromatic ring may optionally be fused or otherwiseattached to other aromatic hydrocarbon rings or non-aromatic hydrocarbonrings. Additionally, the aryl group may be substituted or unsubstitutedby various groups such as hydrogen, halo, hydroxy, alkyl, haloalkyl,alkoxy, nitro, cyano, alkylamine, carboxy or alkoxycarbonyl. Examples ofaryl groups include, for example, phenyl, naphthyl,1,2,3,4-tetrahydronaphthalene, benzodioxole, and biphenyl. Preferredexamples of unsubstituted aryl groups include phenyl and biphenyl.Preferred aryl group substituents include hydrogen, halo, alkyl,haloalkyl, hydroxy and alkoxy.

The term “heteroalkyl”, as used herein alone or as part of a largermoiety, refers to an alkyl radical as defined herein with one or moreheteroatoms replacing a carbon atom with the moiety. Such heteroalkylgroups are alternately referred to using the terms ether, thioether,amine, and the like.

The term “heterocyclyl”, as used herein alone or as part of a largermoiety, refers to saturated, partially unsaturated and unsaturatedheteroatom-containing ring shaped radicals, where the heteroatoms may beselected from nitrogen, sulfur and oxygen. Said heterocyclyl groups maybe unsubstituted or substituted at one or more atoms within the ringsystem. The heterocyclic ring may contain one or more oxo groups.

The term “heterocycloalkyl”, as used herein alone or as part of a largermoiety, refers to a non-aromatic ring system containing at least oneheteroatom selected from nitrogen, oxygen, and sulfur. Theheterocycloalkyl ring may be optionally fused to or otherwise attachedto other heterocycloalkyl rings and/or non-aromatic hydrocarbon rings.Preferred heterocycloalkyl groups have from 3 to 7 members. Examples ofheterocycloalkyl groups include, for example, piperazine, morpholine,piperidine, tetrahydrofuran, pyrrolidine, and pyrazole. Preferredmonocyclic heterocycloalkyl groups include piperidyl, piperazinyl,morpholinyl, pyrrolidinyl, thiomorpholinyl, thiazolidinyl,1,3-dioxolanyl, 1,4-dioxanyl, tetrahydrofuranyl, tetrahydrothiophenyl,tetrahydrothiopyranyl, and the like. Heterocycloalkyl radicals may alsobe partially unsaturated. Examples of such groups includedihydrothienyl, dihydropyranyl, dihydrofuryl, and dihydrothiazolyl.

The term “heteroaryl”, as used herein alone or as part of a largermoiety, refers to an aromatic ring system containing at least oneheteroatom selected from nitrogen, oxygen, and sulfur. The heteroarylring may be fused or otherwise attached to one or more heteroaryl rings,aromatic or non-aromatic hydrocarbon rings or heterocycloalkyl rings.Additionally, the heteroaryl group may be unsubstituted or substitutedat one or more atoms of the ring system, or may contain one or more oxogroups. Examples of heteroaryl groups include, for example, pyridine,furan, thiophene, carbazole and pyrimidine. Preferred examples ofheteroaryl groups include thienyl, benzothienyl, pyridyl, quinolyl,pyrazinyl, pyrimidyl, imidazolyl, benzimidazolyl, furanyl, benzofuranyl,thiazolyl, benzothiazolyl, isoxazolyl, oxadiazolyl, isothiazolyl,benzisothiazolyl, triazolyl, tetrazolyl, pyrrolyl, indolyl, pyrazolyl,benzopyrazolyl, purinyl, benzooxazolyl, and carbazolyl.

The tem “acyl” means an H—C(O)— or alkyl-C(O)— group in which the alkylgroup, straight chain, branched or cyclic, is as previously described.Exemplary acyl groups include formyl, acetyl, propanoyl,2-methylpropanoyl, butanoyl, and caproyl.

The term “aroyl” means an aryl-C(O)— group in which the aryl group is aspreviously described. Exemplary aroyl groups include benzoyl and 1- and2-naphthoyl.

The term “solvate” means a physical association of a compound of thisinvention with one or more solvent molecules. This physical associationinvolves varying degrees of ionic and covalent bonding, includinghydrogen bonding. In certain instances, the solvate will be capable ofisolation, for example when one or more solvent molecules areincorporated in the crystal lattice of the crystalline solid. “Solvate”encompasses both solution-phase and isolatable solvates. Exemplarysolvates include ethanolates, methanolates, and the like. “Hydrate” is asolvate wherein the solvent molecule(s) is/are H₂O.

Examples of aryladamantane compound of the present invention aregenerally represented by Formula 7, shown below:

and pharmaceutically acceptable salts thereof, whereinL is a bond or is —C—(R₃,R₄)—;X is —(R₃,R₄)N(R₅)—, —C(O)N(R₄)—, —N(R₄)C(O)—, —C(R₄,R₅)—, —N(R₄)—, —O—,—S—, —C(O)—, —S(O)₂—, —S(O)₂N(R₄)—, or —N(R₄)S(O)₂—;R₁ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl,heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl,alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl,halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, —COOH,—OH, —SH, —S-alkyl, —CN, —NO₂, —NH₂, —CO₂(alkyl), —OC(O)alkyl,carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, monoor dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl,or mono or dialkylthiocarbamoyl;R₂ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl,heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl,alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl,halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, —COOH,—OH, —SH, —S-alkyl, —CN, —NO₂, —NH₂, —CO₂(alkyl), —OC(O)alkyl,carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, monoor dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl,mono or dialkylthiocarbamoyl, alkyl-S-alkyl, -heteroaryl-aryl,-alkyl-heteroaryl-aryl, —C(O)—NH-aryl, -alkenyl-heteroaryl,—C(O)-heteroaryl, or -alkenyl-heteroaryl-aryl;R₃ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl,heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl,alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl,halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, oxo(═O), —COOH, —OH, —SH, —S-alkyl, —CN, —NO₂, —NH₂, —CO₂(alkyl),—OC(O)alkyl, carbamoyl, mono or dialkylaminocarbamoyl, mono ordialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- ordialkylaminoalkyl, thiocarbamoyl, or mono or dialkylthiocarbamoyl;wherein the alkyl and ring portion of each of the above R₁, R₂, and R₃groups is optionally substituted with up to 5 groups that areindependently (C₁-C₆) alkyl, halogen, haloalkyl, —OC(O)(C₁-C₆ alkyl),—C(O)O(C₁-C₆ alkyl), —CONR′R″, —OC(O)NR′R″, —NR′C(O)R″, —CF₃, —OCF₃,—OH, C₁-C₆ alkoxy, hydroxyalkyl, —CN, —CO₂H, —SH, —S-alkyl, —SOR′R″,—SO₂R′, —NO₂, or NR′R″, wherein R′ and R″ are independently H or (C₁-C₆)alkyl, and wherein each alkyl portion of a substituent is optionallyfurther substituted with 1, 2, or 3 groups independently selected fromhalogen, CN, OH, and NH₂; andR₄ and R₅ are independently H or alkyl, provided that when R₃ and R₄ areon the same carbon and R₃ is oxo, then R₄ is absent.

Aryladamantane compounds of Formula 7 include compounds of formula I-1:

and pharmaceutically acceptable salts thereof, wherein:R₁ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl,heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl,alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl,halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, —COOH,—OH, —SH, —S-alkyl, —CN, —NO₂, —NH₂, —CO₂(alkyl), —OC(O)alkyl,carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, monoor dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl,or mono or dialkylthiocarbamoyl; andR₂ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl,heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl,alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl,halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, —COOH,—OH, —SH, —S-alkyl, —CN, —NO₂, —NH₂, —CO₂(alkyl), —OC(O)alkyl,carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, monoor dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl,mono or dialkylthiocarbamoyl, alkyl-S-alkyl, -heteroaryl-aryl,-alkyl-heteroaryl-aryl, —NH-aryl, -alkenyl-heteroaryl, -heteroaryl,—NH-alkyl, —NH— cycloalkyl, or -alkenyl-heteroaryl-aryl,wherein the alkyl and ring portion of each of the above R₁, and R₂groups is optionally substituted with up to 5 groups that areindependently (C₁-C₆) alkyl, halogen, haloalkyl, —OC(O)(C₁-C₆ alkyl),—C(O)O(C₁-C₆ alkyl), —CONR′R″, —OC(O)NR′R″, —NR′C(O)R″, —CF₃, —OCF₃,—OH, C₁-C₆ alkoxy, hydroxyalkyl, —CN, —CO₂H, —SH, —S-alkyl, —SOR′R″,—SO₂R′, —NO₂, or NR′R″, wherein R′ and R″ are independently H or (C₁-C₆)alkyl, and wherein each alkyl portion of a substituent is optionallyfurther substituted with 1, 2, or 3 groups independently selected fromhalogen, CN, OH, NH₂.

Aryladamantane compounds of Formula 7 include those of formula II:

and pharmaceutically acceptable salts thereof, wherein:Y is —C(R₄,R₅)—, —N(R₄)—, —O—, or —C(O)—;R₁ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl,heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl,alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl,halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, —COOH,—OH, —SH, —S-alkyl, —CN, —NO₂, —NH₂, —CO₂(alkyl), —OC(O)alkyl,carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, monoor dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl,or mono or dialkylthiocarbamoyl;R₂ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl,heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl,alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl,halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, —COOH,—OH, —SH, —S-alkyl, —CN, —NO₂, —NH₂, —CO₂(alkyl), —OC(O)alkyl,carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, monoor dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl,mono or dialkylthiocarbamoyl, alkyl-S-alkyl, -heteroaryl-aryl,-alkyl-heteroaryl-aryl, —C(O)—NH-aryl, -alkenyl-heteroaryl,—C(O)-heteroaryl, or -alkenyl-heteroaryl-aryl;R₃ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl,heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl,alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl,halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, oxo(═O), —COOH, —OH, —SH, —S-alkyl, —CN, —NO₂, —NH₂, —CO₂(alkyl),—OC(O)alkyl, carbamoyl, mono or dialkylaminocarbamoyl, mono ordialkylcarbamoyl, mono or dialkylamino, aminoalkyl, mono- ordialkylaminoalkyl, thiocarbamoyl, or mono or dialkylthiocarbamoyl;wherein the alkyl and ring portion of each of the above R₁, R₂, and R₃groups is optionally substituted with up to 5 groups that areindependently (C₁-C₆) alkyl, halogen, haloalkyl, —OC(O)(C₁-C₆ alkyl),—C(O)O(C₁-C₆ alkyl), —CONR′R″, —OC(O)NR′R″, —NR′C(O)R″, —CF₃, —OCF₃,—OH, C₁-C₆ alkoxy, hydroxyalkyl, —CN, —CO₂H, —SH, —S-alkyl, —SOR′R″,—SO₂R′, —NO₂, or NR′R″, wherein R′ and R″ are independently H or (C₁-C₆)alkyl, and wherein each alkyl portion of a substituent is optionallyfurther substituted with 1, 2, or 3 groups independently selected fromhalogen, CN, OH, NH₂; andR₄ and R₅ are independently H or alkyl.

Compounds of the formula II include those wherein:

Y is —C(R₄,R₅)— or —N(R₄)—;R₁ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl,heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl,alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl,halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, —COOH,—OH, —SH, —S-alkyl, —CN, —NO₂, —NH₂, —CO₂(alkyl), —OC(O)alkyl,carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, monoor dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl,or mono or dialkylthiocarbamoyl;R₂ is H, alkyl, cycloalkyl, cycloalkylalkyl, alkenyl, alkynyl,heteroalkyl, aryl, alkylaryl, alkenylaryl, heterocyclyl, heteroaryl,alkylheteroaryl, heterocycloalkyl, alkyl-heterocycloalkyl, acyl, aroyl,halogen, haloalkyl, alkoxy, haloalkoxy, hydroxyalkyl, alkanoyl, —COOH,—OH, —SH, —S-alkyl, —CN, —NO₂, —NH₂, —CO₂(alkyl), —OC(O)alkyl,carbamoyl, mono or dialkylaminocarbamoyl, mono or dialkylcarbamoyl, monoor dialkylamino, aminoalkyl, mono- or dialkylaminoalkyl, thiocarbamoyl,mono or dialkylthiocarbamoyl, alkyl-S-alkyl, -heteroaryl-aryl,-alkyl-heteroaryl-aryl, —C(O)—NH-aryl, -alkenyl-heteroaryl,—C(O)-heteroaryl, or -alkenyl-heteroaryl-aryl;wherein the alkyl and ring portion of each of the above R₁ and R₂ groupsis optionally substituted with up to 5 groups that are independently(C₁-C₆) alkyl, halogen, haloalkyl, —OC(O)(C₁-C₆ alkyl), —C(O)O(C₁-C₆alkyl), —CONR₄R₅, —OC(O)NR₄R₅, —NR₄C(O)R₅, —CF₃, —OCF₃, —OH, C₁-C₆alkoxy, hydroxyalkyl, —CN, —CO₂H, —SH, —S-alkyl, —SOR₄R₅, —SO₂R₄R₅,—NO₂, or NR₄R₅, and wherein each alkyl portion of a substituent isoptionally further substituted with 1, 2, or 3 groups independentlyselected from halogen, CN, OH, NH₂;R₃ is H, alkyl, or oxo (═O); andR₄ and R₅ are independently H or (C₁-C₆)alkyl.

Representative formula II compounds include:

Cmpd Chemical name Y R3 R1 R2  1 3-(4-Chloro-phenyl)-adamantane-1-carboxylic acidisopropylamide NH ═O

 2 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acidcyclopropylamide NH═O

 3 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(2-ethylsulfanyl-ethyl)- amide NH ═O

 4 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acidphenylamide NH ═O

 5 Adamantane-1-carboxylic acid(4-hydroxy-phenyl)- amide NH ═O H

 6 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid(4-hydroxy-phenyl)-amide NH ═O

 7 Acetic acid 4-{[3-(4-chloro- phenyl)-adamantane-1-carbonyl]-amino}-phenyl ester NH ═O

 8 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(2,4-dihydroxy-phenyl)- amide NH ═O

 9 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid(3-hydroxymethyl-phenyl)-amide NH ═O

 10 Adamantane-1-carboxylic acid(4-cyanomethyl- phenyl)-amide NH ═O H

 11 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid(4-cyanomethyl-phenyl)-amide NH ═O

 12 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acidbenzylamide NH ═O

 13 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid4-tert-butyl-benzylamide NH ═O

 14 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid4-methylsulfanyl-benzylamide NH ═O

 15 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid3-trifluoromethyl-benzylamide NH ═O

 16 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid4-trifluoromethyl-benzylamide NH ═O

 17 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid3,5-bis-trifluoromethyl- benzylamide NH ═O

 18 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid3-fluoro-5-trifluoromethyl- benzylamide NH ═O

 19 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid2-fluoro-4-trifluoromethyl- benzylamide NH ═O

 20 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid3,5-difluoro-benzylamide NH ═O

 21 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid3,4-difluoro-benzylamide NH ═O

 22 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid3,4,5-trifluoro-benzylamide NH ═O

 23 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid3-chloro-4-fluoro-benzylamide NH ═O

 24 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid4-fluoro-3-trifluoromethyl- benzylamide NH ═O

 25 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid2-chloro-4-fluoro-benzylamide NH ═O

 26 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid4-chloro-3-trifluoromethyl- benzylamide NH ═O

 27 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid3-aminomethyl-2,4,5,6- tetrachloro-benzylamide NH ═O

 28 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid[1-(4-chloro-phenyl)- ethyl]-amide NH ═O

 29 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid[1-(4-bromo-phenyl)- ethyl]-amide NH ═O

 30 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid4-methanesulfonyl-benzylamide NH ═O

 31 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid4-dimethylamino-benzylamide NH ═O

 32 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid4-trifluoromethoxy-benzylamide NH ═O

 33 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid3-trifluoromethoxy-benzylamide NH ═O

 34 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid4-phenoxy-benzylamide NH ═O

 35 Adamantane-1-carboxylic acid3,4-dihydroxy- benzylamide NH ═O H

 36 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid3,4-dihydroxy-benzylamide NH ═O

 37 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acidphenethyl-amide NH═O

 38 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid[2-(4-fluoro-phenyl)- ethyl]-amide NH ═O

 39 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid[2-(4-bromo-phenyl)- ethyl]-amide NH ═O

 40 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid[2-(4-hydroxy-phenyl)- ethyl]-amide NH ═O

 41 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid4-phenoxy-benzylamide NH ═O

 42 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid[2-(3-bromo-4-methoxy-phenyl)-ethyl]- amide NH ═O

 43 Adamantane-1-carboxylic acid[2-(3,4-dihydroxy- phenyl)-ethyl]-amideNH ═O H

 44 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid[2-(3,4-dihydroxy-phenyl)-ethyl]-amide NH ═O

 45 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(2-benzo[1,3]dioxol-5- yl-ethyl)-amide NH ═O

 46 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid[2-(3-phenoxy-phenyl)- ethyl]-amide NH ═O

 47 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid[2-(4-phenoxy-phenyl)- ethyl]-amide NH ═O

 48 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid(3-phenyl-propyl)-amide NH ═O

 49 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(biphenyl-4-ylmethyl)- amide NH ═O

 50 Adamantane-1-carboxylic acid(1-methyl-piperidin-4- yl)-amide NH ═O H

 51 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(1-methyl-piperidin-4- yl)-amide NH ═O

 52 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(4-methyl-piperazin-1- yl)-amide NH ═O

 53 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid(3-tert-butylamino-propyl)-amide NH ═O

 54 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid(3-pyrrolidin-1-yl-propyl)-amide NH ═O

 55 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid[3-(2-oxo-pyrrolidin-1- yl)-propyl]-amide NH ═O

 56 Adamantane-1-carboxylic acid[2-(1-methyl-pyrrolidin-2-yl)-ethyl]-amide NH ═O H

 57 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid[2-(1-methyl-pyrrolidin- 2-yl)-ethyl]-amide NH ═O

 58 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid(2-morpholin-4-yl-ethyl)-amide NH ═O

 59 3-(4-Chloro-phenyl)- adaniantane-1-carboxylicacid(2-piperazin-1-yl-ethyl) amide NH ═O

 60 Adamantane-1-carboxylic acid(pyridin-4-ylmethyl)- amide NH ═O H

 61 3-(4-Fluoro-phenyl)- adamantane-1-carboxylicacid(pyridin-4-ylmethyl)- amide NH ═O

 62 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(pyridin-4-ylmethyl)- amide NH ═O

 63 Adamantane-1-carboxylic acid(pyridin-4-ylmethyl)- amide NH ═O H

 64 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(2-pyridin-4-yl-ethyl)- amide NH ═O

 65 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid(3-imidazol-1-yl-propyl)-amide NH ═O

 66 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(2-methyl-1H-indol-5- yl)-amide NH ═O

 67 3-(4-Chloro-phenyl)- adamantane-1-carboxylic acid(1H-tetrazol-5-yl)-amide NH ═O

 68 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(9-ethyl-9H-carbazol-3- yl)-amide NH ═O

 69 Adamantane-1-carboxylic acid[4-(4-chloro-phenyl)-thiazol-2-yl]-amide NH ═O H

 70 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid[4-(4-chloro-phenyl)- thiazol-2-yl]-amide NH ═O

 71 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacidbenzothiazol-2-ylamide NH ═O

 72 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(5-chloro-benzooxazol- 2-yl)-amide NH ═O

 73 3-(4-Chloro-phenyl)- adamantane-1-carboxylicacid(9H-purin-6-yl)-amide NH ═O

 75 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]- isopropyl-amine NH H

 76 4- and -phenol NH H

 77 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-(4-trifluoromethyl-benzyl)- amine NH H

 78 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-(2-fluoro-4-trifluoromethyl- benzyl)-amine NH H

 79 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-(4-fluoro-3-trifluoromethyl- benzyl)-amine NH H

 80 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-(4-trifluoromethoxy-benzyl)- amine NH H

 81 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-[2-(3-phenoxy-phenyl)-ethyl]- amine NH H

 82 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-(1-methyl-piperidin-4-yl)- amine NH H

 83 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-(4-methyl-piperazin-1-yl)- amine NH H

 84 N-tert-Butyl-N′-[3-(4- chloro-phenyl)-adamantan-1-ylmethyl]-propane-1,3- diamine NH H

 85 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-(3-pyrrolidin-1-yl-propyl)- amine NH H

 86 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-[2-(1-methyl-pyrrolidin-2-yl)- ethyl]-amine NH H

 87 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-(2-morpholin-4-yl-ethyl)- amine NH H

 88 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-pyridin-4-ylmethyl-amine NH H

 89 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-(9-ethyl-9H-carbazol-3-yl)- amine NH H

 90 [3-(4-Chloro-phenyl)- adamantan-1-ylmethyl]-[5-(4-chloro-phenyl)-thiazol-2- yl]-amine NH H

 91 1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethylamine NH CH3

H  92 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}- isopropyl-amineNH CH3

 93 Phenyl-[1-(3-phenyl- adamantan-1-yl)-ethyl]- amine NH CH3

 94 {1-[3-(4-Fluoro-phenyl)- adamantan-1-yl]-ethyl}- phenyl-amine NH CH3

 95 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}- phenyl-amine NH CH3

 96 (1-Adamantan-1-yl-ethyl)- benzyl-amine NH CH3 H

 97 Benzyl-[1-(3-phenyl- adamantan-1-yl)-ethyl]- amine NH CH3

 98 Benzyl-{1-[3-(4-fluoro- phenyl)-adamantan-1-yl]- ethyl}-amine NH CH3

 99 Benzyl-{1-[3-(4-chloro- phenyl)-adamantan-1-yl]- ethyl}-amine NH CH3

100 (4-tert-Butyl-benzyl)-{1-[3- (4-chloro-phenyl)-adamantan-1-yl]-ethyl}- amine NH CH3

101 [1-(4-Bromo-phenyl)-ethyl]- {1-[3-(4-chloro-phenyl)-adamantan-1-yl]-ethyl}- amine NH CH3

102 (1-Adamantan-1-yl-ethyl)- [2-(4-bromo-phenyl)-ethyl]- amine NH CH3 H

103 [2-(4-Bromo-phenyl)-ethyl]- {1-[3-(4-chloro-phenyl)-adamantan-1-yl]-ethyl}- amine NH CH3

104 (1-Adamantan-1-yl-ethyl)- (1-methyl-piperidin-4-yl)- amine NH CH3 H

105 (1-Methyl-piperidin-4-yl)- [1-(3-phenyl-adamantan-1-yl)-ethyl]-amine NH CH3

106 {1-[3-(4-Fluoro-phenyl)- adamantan-1-yl]-ethyl}-(1-methyl-piperidin-4-yl)- amine NH CH3

107 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-(1-methyl-piperidin-4-yl)- amine NH CH3

108 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-(4-methyl-piperazin-1-yl)- amine NH CH3

109 {1-[3-(Phenyl)-adamantan- 1-yl]-ethyl}-pyridin-4- ylmethyl-amine NHCH3

110 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-(6-chloro-pyridin-3-ylmethyl)- amine NH CH3

111 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-(2-pyridin-4-yl-ethyl)-amine NH CH3

112 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-(3H-imidazol-4-ylmethyl)- amine NH CH3

113 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-(2-methyl-1H-indol-5-yl)- amine NH CH3

114 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-(9-ethyl-9H-carbazol-3-yl)- amine NH CH3

115 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-(9-ethyl-9H-carbazol-3- ylmethyl)-amine NH CH3

116 9-Ethyl-9H-carbazole-3- carboxylic acid {1-[3-(4-chloro-phenyl)-adamantan- 1-yl]-ethyl}-amide NH CH3

117 1-{1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-3-(4-chloro-3-trifluoromethyl- phenyl)-urea NH CH3

118 1-{1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-3-(4-chloro-3-trifluoromethyl- phenyl)-urea NH CH3

119 (4-Bromo-thiophen-2- ylmethyl)-{1-[3-(4-chloro-phenyl)-adamantan-1-yl]- ethyl}-amine NH CH3

120 {1-[3-(4-Chloro-phenyl)- adamantan-1-yl]-ethyl}-(4-phenyl-thiophen-2- ylmethyl)-amine NH CH3

Representative formula I-1 compounds include:

Cmpd Chemical name R1 R2 121 3-Phenyl-adamantane-1-carboxylicacid

OH 122 3-(4-Fluoro-phenyl)-adamantane-1- carboxylic acid

OH 123 3-(4-Chloro-phenyl)-adamantane-1- carboxylic acid

OH 124 1-Adamantan-1-yl-ethanone H CH3 1251-(3-Phenyl-adamantan-1-yl)-ethanone

CH3 126 1-[3-(4-Fluoro-phenyl)-adamantan-1-yl]- ethanone

CH3 127 1-[3-(4-Chloro-phenyl)-adamantan-1-yl]- ethanone

CH3 128 2-(Adamantane-1-carbonyl)-malonicacid dimethyl ester H

129 2-[3-(4-Chloro-phenyl)-adamantane-1- carbonyl]-malonic aciddimethylester

130 3-(4-Chloro-phenyl)-1-[3-(4-chloro-phenyl)-adamantan-1-yl]-propenone

131 4-{3-[3-(4-Chloro-phenyl)-adamantan-1-yl]-3-oxo-propenyl}-benzonitrile

132 1-[3-(4-Chloro-phenyl)-adamantan-1-yl]-3-(4-hydroxy-phenyl)-propenone

133 1-[3-(4-Chloro-phenyl)-adamantan-1-yl]- 3-naphthalen-2-yl-propenone

134 1-[3-(4-Chloro-phenyl)-adamantan-1-yl]-3-(6-chloro-pyridin-3-yl)-propenone

135 1-[3-(4-Chloro-phenyl)-adamantan-1-yl]-3-(1H-imidazol-4-yl)-propenone

136 1-[3-(4-Chloro-phenyl)-adamantan-1-yl]-3-(9-ethyl-9H-carbazol-3-yl)-propenone

137 1-[3-(4-Chloro-phenyl)-adamantan-1-yl]-3-(4-phenyl-thiophen-2-yl)-propenone

A particularly preferred aryladamantane compound of the presentinvention is illustrated below and referred to as ABC294640[3-(4-chlorophenyl)-adamantane-1-carboxylic acid(pyridin-4-ylmethyl)amide]:

In an embodiment, an aryladamantane compound of the present invention isselected from a compound of Formula 8:

and pharmaceutically acceptable salts thereof, wherein

-   -   R₁ is H, Cl or F;    -   R₂ is H or alkyl;    -   m is 0, 1 or 2;    -   n is 1, 2, 3, 4 or 5;    -   each R₃ is independently H, —C(O)alkyl, —C(O)CH₂CH₂C(O)OH, R₄,        —C(O)NR₅R₆, —P(O)(OR₇)₂ or glucosyl, provided that at least one        R₃ is not H,    -   wherein        -   R₄ is a natural or unnatural amino acid linked through the            carboxyl moiety as an ester,        -   R₅ is H or alkyl,        -   R₆ is H or alkyl, and

each R₇ is independently H or alkyl.

In certain embodiments of the compounds of formula (I) as describedabove, the

moiety is a catechol with substitution at least one catechol —OH. Forexample, in one embodiment, the

moiety has the structure

In one particularly preferred embodiment of the compounds of formula (I)as described above, the

moiety has the structure

In one especially preferred embodiment of the invention, compounds offormula (I) have R₁=Cl, R₂=H, m=2, n=2, and each R₃=—C(O)alkyl,especially —C(O)CH₃.

For example, compounds of the invention include:

-   Acetic acid    2-acetoxy-5-(2-{[3-(4-chlorophenyl)-adamantane-1-carbonyl]-amino}ethyl)phenyl    ester;-   Propionic acid    2-propionyloxy-5-(2-{[3-(4-chlorophenyl)-adamantane-1-carbonyl]-amino}ethyl)phenyl    ester;-   Butyric acid    2-butyryloxy-5-(2-{[3-(4-chlorophenyl)-adamantane-1-carbonyl]-amino}ethyl)phenyl    ester;-   Isobutyric acid    5-(2-([3-(4-chlorophenyl)adamantane-1-carbonyl]amino)ethyl)-2-hydroxyphenyl    ester; and-   2-Amino-3-methyl-butyric acid    5-(2-{[3-(4-chlorophenyl)adamantane-1-carbonyl]amino}ethyl)-2-hydroxyphenyl    ester.

A particularly preferred aryladamantane compound of the presentinvention is illustrated below and referred to as ABC294735[3-(4-chlorophenyl)adamantane-1-carboxylic acid[2-(3,4-dihydroxyphenyl)ethyl]amide]:

Solid forms for oral administration may contain pharmaceuticallyacceptable binders, sweeteners, disintegrating agents, diluents,flavorings, coating agents, preservatives, lubricants, and/or time delayagents. Suitable binders include gum acacia, gelatin, corn starch, gumtragacanth, sodium alginate, carboxymethylcellulose or polyethyleneglycol (PEG). Suitable sweeteners include sucrose, lactose, glucose,aspartame or saccharine. Suitable disintegrating agents include cornstarch, methylcellulose, polyvinylpyrrolidone, xanthan gum, bentonite,alginic acid or agar. Suitable diluents include lactose, sorbitol,mannitol, dextrose, kaolin, cellulose, calcium carbonate, calciumsilicate or dicalcium phosphate. Suitable flavoring agents includepeppermint oil, oil of wintergreen, cherry, orange, or raspberryflavoring. Suitable coating agents include polymers or copolymers ofacrylic acid and/or methacrylic acid and/or their esters, waxes, fattyalcohols, zein, shellac or gluten. Suitable preservatives include sodiumbenzoate, vitamin E, alpha-tocopherol, ascorbic acid, methyl paraben,propyl paraben or sodium bisulphite. Suitable lubricants includemagnesium stearate, stearic acid, sodium oleate, sodium chloride ortalc. Suitable time delay agents include glyceryl monostearate orglyceryl distearate.

In an embodiment, the present disclosure relates to methods for treatinga subject having a ssRNA viral infection, such as Ebola virus or Marburgvirus, by concomitantly administering i) a therapeutically effectiveamount of an anti-viral drug; and ii) a therapeutically effective amountof at least one anti-atypical mycobacterial agent. In an embodiment, theanti-viral drug is Brivudine (BVDU), an active metabolite of BVDU, asalt thereof, or BVDU in protected or in prodrug form. Suitableanti-atypical mycobacterial agents include, but are not limited to,clarithromycin, rifabutin, rifampicin, azithromycin, roxithromycin,amikacin, clofazimine, ethambutol ofloxacin, ciprofloxacin andoxazolidinone. In an embodiment, the anti-atypical mycobacterial agentsare selected from at least one of rifabutin, clarithromycin andclofazimine. In an embodiment, the anti-atypical mycobacterial agent isclofazimine. In an embodiment, the anti-atypical mycobacterial agentclofazimine is administered as a single solid oral dosage form.

In an embodiment, the present disclosure relates to methods for treatinga subject having a ssRNA viral infection, such as Ebola virus, byconcomitantly administering i) a therapeutically effective amount of ananti-viral drug; ii) a therapeutically effective amount of rifabutin;iii) a therapeutically effective amount of clofazimine; and iv) atherapeutically effective amount of clarithromycin. In an embodiment,the anti-viral drug is Brivudine (BVDU), an active metabolite of BVDU, asalt thereof, or BVDU in protected or in prodrug form.

In an embodiment, the present disclosure relates to methods for treatinga subject having a ssRNA viral infection, e.g., but not limited to,Ebola virus, by concomitantly administering i) a therapeuticallyeffective amount of an anti-viral and ii) a therapeutically effectiveamount of clofazamine. In an embodiment, the anti-viral drug isBrivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDUin protected or in prodrug form.

In an embodiment, the present disclosure relates to methods for treatinga subject having a ssRNA viral infection, e.g., but not limited to,Ebola virus, by concomitantly administering i) a therapeuticallyeffective amount of an anti-viral drug; ii) a therapeutically effectiveamount of rifabutin; and iii) a therapeutically effective amount ofclofazimine. In an embodiment, the anti-viral drug is Brivudine (BVDU),an active metabolite of BVDU, a salt thereof, or BVDU in protected or inprodrug form. In an embodiment, the rifabutin and the clofazimine areadministered as a single solid oral dosage form. In an embodiment,rifabutin and clofazimine are administered as separate solid oral dosageforms.

In an embodiment, the present disclosure relates to methods for treatinga subject having a ssRNA viral infection, e.g., but not limited to,Ebola virus, by concomitantly administering i) a therapeuticallyeffective amount of brivudine, an active metabolite of brivudine (BVDU),an active metabolite of BVDU, a salt thereof, or BVDU in protected or inprodrug form and ii) a therapeutically effective amount ofclarithromycin as an intravenous infusion.

In an embodiment, the present disclosure relates to methods for treatinga subject having a ssRNA viral infection, e.g., but not limited to,Ebola virus, by administering a therapeutically effective amount of anaryladamantane compound. In an embodiment the aryladamantane compound isselected from a compound of formula 7.

In an embodiment, the present disclosure relates to methods for treatinga subject having a ssRNA viral infection, e.g., but not limited to,Ebola virus, by concomitantly administering i) a therapeuticallyeffective amount of brivudine, an active metabolite of brivudine (BVDU),an active metabolite of BVDU, a salt thereof, or BVDU in protected or inprodrug form and ii) a therapeutically effective amount of anaryladamantane compound. In an embodiment the aryladamantane compound isselected from a compound of formula 7.

In an embodiment, the present disclosure relates to methods for treatinga subject having a ssRNA viral infection, such as Ebola virus or Marburgvirus, by concomitantly administering i) a therapeutically effectiveamount of an anti-viral drug; ii) a therapeutically effective amount ofat least one anti-atypical mycobacterial agent; and iii) atherapeutically effective amount of an aryladamantane compound. In anembodiment, the anti-viral drug is Brivudine (BVDU), an activemetabolite of BVDU, a salt thereof, or BVDU in protected or in prodrugform. Suitable anti-atypical mycobacterial agents include, but are notlimited to, clarithromycin, rifabutin, rifampicin, azithromycin,roxithromycin, amikacin, clofazimine, ethambutol ofloxacin,ciprofloxacin and oxazolidinone. In an embodiment, the anti-atypicalmycobacterial agents are selected from at least one of rifabutin,clarithromycin and clofazimine. In an embodiment, the anti-atypicalmycobacterial agent is clofazimine. In an embodiment, the anti-atypicalmycobacterial agent clofazimine is administered as a single solid oraldosage form. In an embodiment the aryladamantane compound is selectedfrom a compound of formula 7.

In an embodiment, a treatment for a ssRNA viral infection, such as Ebolavirus includes providing intravenous fluids (IV) and balancingelectrolytes (body salts) to a subject; maintaining oxygen status andblood pressure of the subject; and concomitantly administering i) atherapeutically effective amount of an anti-viral drug; ii) atherapeutically effective amount of rifabutin; iii) a therapeuticallyeffective amount of clofazimine; and iv) a therapeutically effectiveamount of clarithromycin. In an embodiment, the anti-viral drug isBrivudine (BVDU), an active metabolite of BVDU, a salt thereof, or BVDUin protected or in prodrug form.

In an embodiment, a treatment for a ssRNA viral infection, e.g., but notlimited to, Ebola virus, includes providing intravenous fluids (IV) andbalancing electrolytes (body salts) to a subject; maintaining oxygenstatus and blood pressure of the subject; and concomitantlyadministering i) a therapeutically effective amount of an anti-viraldrug; and ii) a therapeutically effective amount of clofazimine. In anembodiment, the anti-viral drug is Brivudine (BVDU), an activemetabolite of BVDU, a salt thereof, or BVDU in protected or in prodrugform.

In an embodiment, a treatment for a ssRNA viral infection, e.g., but notlimited to, Ebola virus, includes providing intravenous fluids (IV) andbalancing electrolytes (body salts) to a subject; maintaining oxygenstatus and blood pressure of the subject; and concomitantlyadministering i) a therapeutically effective amount of an anti-viraldrug; ii) a therapeutically effective amount of clofazimine; and iii) atherapeutically effective amount of rifabutin. In an embodiment, theanti-viral drug is Brivudine (BVDU), an active metabolite of BVDU, asalt thereof, or BVDU in protected or in prodrug form. In an embodiment,the rifabutin and the clofazimine are administered as a single solidoral dosage form. In an embodiment, rifabutin and clofazimine areadministered as separate solid oral dosage forms.

In an embodiment, a treatment for a ssRNA viral infection, e.g., but notlimited to, Ebola virus, includes providing intravenous fluids (IV) andbalancing electrolytes (body salts) to a subject; maintaining oxygenstatus and blood pressure of the subject; and concomitantlyadministering i) a therapeutically effective amount of an anti-viraldrug; and ii) a therapeutically effective amount of clarithromycin as anintravenous infusion. In an embodiment, the anti-viral drug is Brivudine(BVDU), an active metabolite of BVDU, a salt thereof, or BVDU inprotected or in prodrug form.

In an embodiment, a treatment for a ssRNA viral infection, e.g., but notlimited to, Ebola virus, includes providing intravenous fluids (IV) andbalancing electrolytes (body salts) to a subject; maintaining oxygenstatus and blood pressure of the subject; and administering atherapeutically effective amount of an aryladamantane compound. In anembodiment the aryladamantane compound is selected from a compound offormula 7.

In an embodiment, a treatment for a ssRNA viral infection, e.g., but notlimited to, Ebola virus, includes providing intravenous fluids (IV) andbalancing electrolytes (body salts) to a subject; maintaining oxygenstatus and blood pressure of the subject; and concomitantlyadministering i) a therapeutically effective amount of an anti-viraldrug; and ii) a therapeutically effective amount of at least oneanti-atypical mycobacterial agent. In an embodiment, the anti-viral drugis Brivudine (BVDU), an active metabolite of BVDU, a salt thereof, orBVDU in protected or in prodrug form. Suitable anti-atypicalmycobacterial agents include, but are not limited to, clarithromycin,rifabutin, rifampicin, azithromycin, roxithromycin, amikacin,clofazimine, ethambutol ofloxacin, ciprofloxacin and oxazolidinone. Inan embodiment, the anti-atypical mycobacterial agents are selected fromat least one of rifabutin, clarithromycin and clofazimine. In anembodiment, the anti-atypical mycobacterial agent is clofazimine. In anembodiment, the anti-atypical mycobacterial agent clofazimine isadministered as a single solid oral dosage form.

In an embodiment, a treatment for a ssRNA viral infection, e.g., but notlimited to, Ebola virus, includes providing intravenous fluids (IV) andbalancing electrolytes (body salts) to a subject; maintaining oxygenstatus and blood pressure of the subject; and concomitantlyadministering i) a therapeutically effective amount of an anti-viraldrug; ii) a therapeutically effective amount of at least oneanti-atypical mycobacterial agent; and iii) a therapeutically effectiveamount of an aryladamantane compound. In an embodiment, the anti-viraldrug is Brivudine (BVDU), an active metabolite of BVDU, a salt thereof,or BVDU in protected or in prodrug form. Suitable anti-atypicalmycobacterial agents include, but are not limited to, clarithromycin,rifabutin, rifampicin, azithromycin, roxithromycin, amikacin,clofazimine, ethambutol ofloxacin, ciprofloxacin and oxazolidinone. Inan embodiment, the anti-atypical mycobacterial agents are selected fromat least one of rifabutin, clarithromycin and clofazimine. In anembodiment, the anti-atypical mycobacterial agent is clofazimine. In anembodiment, the anti-atypical mycobacterial agent clofazimine isadministered as a single solid oral dosage form. In an embodiment thearyladamantane compound is selected from a compound of formula 7.

If desired, the compounds of the invention may be employed inmechanistic assays to determine whether other combinations, or singleagents, are as effective as the combinations of the invention ininhibiting a viral disease (e.g., those described herein) using assaysgenerally known in the art. For example, candidate compounds may betested, alone or in combination (e.g., with an agent that inhibits viralreplication, such as those described herein) and applied to cells (e.g.,hepatic cells such as HepG2, kidney epithelial cells such as 293T,macrophages such as THP-1, or isolated primary cells). After a suitabletime, viral replication or load of these cells is examined. A decreasein viral replication or viral load identifies a candidate compound orcombination of agents as an effective agent for treating a viraldisease.

The compositions and methods of the invention can include formulation(s)of compound(s) that, upon administration to a subject, result in aconcentration of the compound(s) that treats a filovirus-mediateddisease. The compound(s) may be contained in any appropriate amount inany suitable carrier substance, and are generally present in an amountof 1-95% by weight of the total weight of the composition. Thecomposition may be provided in a dosage form that is suitable for theoral, parenteral (e.g., intravenously or intramuscularly), rectal,dermatological, cutaneous, nasal, vaginal, inhalant, skin (patch),ocular, intrathecal, or intracranial administration route. Thus, thecomposition may be in the form of, e.g., tablets, capsules, pills,powders, granulates, suspensions, emulsions, solutions, gels includinghydrogels, pastes, ointments, creams, plasters, drenches, osmoticdelivery devices, suppositories, enemas, injectables, implants, sprays,or aerosols. The pharmaceutical compositions may be formulated accordingto conventional pharmaceutical practice.

Pharmaceutical compositions according to the invention or used in themethods of the invention may be formulated to release the activecompound immediately upon administration or at any predetermined time ortime period after administration. The latter types of compositions aregenerally known as controlled release formulations, which include (i)formulations that create substantially constant concentrations of theagent(s) of the invention within the body over an extended period oftime; (ii) formulations that after a predetermined lag time createsubstantially constant concentrations of the agent(s) of the inventionwithin the body over an extended period of time; (iii) formulations thatsustain the agent(s) action during a predetermined time period bymaintaining a relatively constant, effective level of the agent(s) inthe body with concomitant minimization of undesirable side effectsassociated with fluctuations in the plasma level of the agent(s)(sawtooth kinetic pattern); (iv) formulations that localize action ofagent(s), e.g., spatial placement of a controlled release compositionadjacent to or in the diseased tissue or organ; (v) formulations thatachieve convenience of dosing, e.g., administering the composition onceper week or once every two weeks; and (vi) formulations that target theaction of the agent(s) by using carriers or chemical derivatives todeliver the combination to a particular target cell type.

Any of a number of strategies can be pursued in order to obtaincontrolled release in which the rate of release outweighs the rate ofmetabolism of the compound in question. In one example, controlledrelease is obtained by appropriate selection of various formulationparameters and ingredients, including, e.g., various types of controlledrelease compositions and coatings. Thus, the compound(s) are formulatedwith appropriate excipients into a pharmaceutical composition that, uponadministration, releases the compound(s) in a controlled manner.Examples include single or multiple unit tablet or capsule compositions,oil solutions, suspensions, emulsions, microcapsules, molecularcomplexes, microspheres, nanoparticles, patches, and liposomes.

It is not intended that administration of compounds be limited to asingle formulation and delivery method for all compounds of acombination. The combination can be administered using separateformulations and/or delivery methods for each compound of thecombination using, for example, any of the above-described formulationsand methods. In one example, a first agent is delivered orally, and asecond agent is delivered intravenously.

The dosage of a compound or a combination of compounds depends onseveral factors, including: the administration method, the type ofdisease to be treated, the severity of the infection, whetheradministration first occurs at an early or late stage of infection, andthe age, weight, and health of the patient to be treated. Forcombinations that include a synergistic pair of agents identifiedherein, the recommended dosage for the anti-viral agent can be less thanor equal to the recommended dose as given in the Physician's DeskReference, 69^(th) Edition (2015).

As described above, the compound(s) in question may be administeredorally in the form of tablets, capsules, elixirs or syrups, or rectallyin the form of suppositories. Parenteral administration of a compound issuitably performed, for example, in the form of saline solutions or withthe compound(s) incorporated into liposomes. In cases where the compoundin itself is not sufficiently soluble to be dissolved, a solubilizersuch as ethanol can be applied. The correct dosage of a compound can bedetermined by examining the efficacy of the compound in viralreplication assays, as well as its toxicity in humans.

The agents of the invention are also useful tools in elucidatingmechanistic information about the biological pathways involved in viraldiseases. Such information can lead to the development of newcombinations or single agents for treating, preventing, or reducing aviral disease. Methods known in the art to determine biological pathwayscan be used to determine the pathway, or network of pathways affected bycontacting cells (e.g., primary macrophage cells) infected with a viruswith the compounds of the invention. Such methods can include, analyzingcellular constituents that are expressed or repressed after contact withthe compounds of the invention as compared to untreated, positive ornegative control compounds, and/or new single agents and combinations,or analyzing some other activity of the cell or virus such as anenzymatic activity, nutrient uptake, and proliferation. Cellularcomponents analyzed can include gene transcripts, and proteinexpression. Suitable methods can include standard biochemistrytechniques, radiolabeling the compounds of the invention (e.g., ¹⁴C or³H labeling), and observing the compounds binding to proteins, e.g.,using 2D gels, gene expression profiling. Once identified, suchcompounds can be used in in vivo models (e.g., knockout or transgenicmice) to further validate the tool or develop new agents or strategiesto treat viral disease.

EXAMPLES

The following examples are intended to illustrate rather than limit theinvention.

Example 1 In Vitro Screening Experiments

A screening experiment, the in-vitro plaque assay, provided resultsregarding measurable inhibition of replication of ssRNA viruses in cellculture.

In order to perform the assay, primary human macrophage cells wereseeded at 1×10⁵ cells/well in 100 μl 10% RPMI 1640 medium (Biocoatcollagen I-coated 96 well plates). The next day, the medium was removedand 100 μl of medium containing different concentrations of drugs wasadded for one (1) hour. Afterwards, 15 μl of virus medium mixture wasadded to each well for one (1) hour (multiplicity of infection (MOI) ofeither 0.01, 0.1 or 1.0). The virus inoculars were removed and 100 μl ofmedium containing different concentrations of drugs was added. Theplates were kept at 37° C. incubator for 48 h. After 48 h postinfection, the supernatants were harvested for plaque assay and theplates were fixed with 10% NBF. 7. The plates were then stained withanti-EBOV VP40. The plates were washed three (3) times with PBS toremove NBF. Cells were permeablized with 0.25% Triton X-100 in PBS for 5minutes at room temperature. The plates were again washed three (3)times with PBS, and then blocked with 10% BSA in PBS for 30 minutes at37° C. The cells were stained with primary antibody (anti-EBOV Vp40BMD004B007) at 1:1000 in 3% BSA (in PBS) for 2 h at 37° C. The plateswere again washed three (3) times with PBS and stained with a secondaryantibody (Alex-488 conjugated goat anti-mouse IgG) at 1:2000 in 3% BSA(in PBS) for 45 minutes at 37° C. The plates were again washed three (3)times with PBS and cells were stained with Horst for 10 minutes at roomtemperature. Plates were washed three (3) times with PBS and thenscanned using the PerkinElmer Operetta® High Content Imaging System. Thepercentage of infected cells was measured and analyzed using theOperetta® High Content Imaging System with Harmony® High Content Imagingand Analysis Software.

Primary human macrophage cells were seeded at 1×10⁵ cells/well in 100 μl10% RPMI 1640 medium (Biocoat collagen I-coated 96 well plates). Thenext day, the medium was removed and 100 μl medium containing differentconcentration of drugs was added. 48 h post treatment, cytotoxicity wasmeasured using Dojindo's Cell Counting Kit-8 (CCK-8), which utilizesDojindo's highly water-soluble tetrazolium salt (WST). WST-8 produces awater-soluble formazan dye upon reduction in the presence of an electronmediator. The viability was calculated with regard to the untreated cellcontrol, which was set to 100% viability.

The assays described above were used to identify drugs that inhibit thereplication of Ebola virus in primary human macrophage cells.

The Table below provides a listing of the concentration of drugs anddrug combinations tested and their effect on primary human macrophagecell toxicity.

Drug(s) Cytotoxic to Primary concentration Human Macrophage Cells?Clofazamine 6 μM No Clofazamine 7 μM No Clofazamine 8 μM No Clofazamine9 μM No Rifabutin 30 μM No Rifabutin 45 μM No Rifabutin 60 μM NoRifabutin 75 μM Yes Rifabutin 90 μM Yes Rifabutin 120 μM Yes Rifabutin150 μM Yes Rifabutin 180 μM Yes Rifabutin 210 μM Yes Rifabutin 240 μMYes Rifabutin 270 μM Yes Rifabutin 300 μM Yes Rifabutin 330 μM YesRifabutin 360 μM Yes Clarithromycin 60 μM No Clarithromycin 75 μM NoClarithromycin 90 μM No Clarithromycin 120 μM No Clarithromycin 150 μMNo Clarithromycin 180 μM No Clarithromycin 210 μM No Clarithromycin 240μM Yes Clarithromycin 270 μM Yes Clarithromycin 300 μM YesClarithromycin 330 μM Yes Clarithromycin 360 μM Yes Brivudine 90 μM NoBrivudine 120 μM No Brivudine 150 μM No Brivudine 180 μM No Brivudine210 μM No Brivudine 240 μM No Brivudine 270 μM No Brivudine 300 μM YesBrivudine 330 μM Yes Brivudine 360 μM Yes ABC29460 2.5 μM No ABC294605.0 μM No ABC29460 10 μM No ABC29460 20 μM Yes ABC29460 40 μM YesABC29460 60 μM Yes ABC29460 80 μM Yes ABC29460 100 μM Yes Clarithromycin210 μM No Brivudine 270 μM Rifabutin 45 μM No Clarithromycin 75 μMRifabutin 45 μM No Clarithromycin 90 μM Rifabutin 45 μM NoClarithromycin 120 μM Rifabutin 45 μM No Clarithromycin 150 μM Rifabutin45 μM No Clarithromycin 180 μM Rifabutin 45 μM No Clarithromycin 210 μMRifabutin 60 μM Yes Clarithromycin 210 μM Rifabutin 60 μM Yes Brivudine270 μM Rifabutin 60 μM Yes Clarithromycin 210 μM Brivudine 270 μMClofazamine 13.2 μM Yes Rifabutin 31.8 μM Clarithromycin 76.2 μMClofazamine 11 μM Yes Rifabutin 26.5 μM Clarithromycin 63.5 μMClofazamine 8.8 μM Yes Rifabutin 21.2 μM Clarithromycin 50.8 μMClofazamine 6.6 μM No Rifabutin 15.9 μM Clarithromycin 38.1 μMClofazamine 4.4 μM No Rifabutin 10.6 μM Clarithromycin 25.4 μMClofazamine 2.2 μM No Rifabutin 5.3 μM Clarithromycin 12.7 μMClofazamine 1.1 μM No Rifabutin 2.65 μM Clarithromycin 6.35 μMClofazamine 0.66 μM No Rifabutin 1.59 μM Clarithromycin 3.81 μMClofazamine 0.22 μM No Rifabutin 0.53 μM Clarithromycin 1.27 μMClofazamine 0.11 μM No Rifabutin 0.265 μM Clarithromycin 0.635 μMClofazamine 13.2 μM Yes Rifabutin 31.8 μM Brivudine 90.0 μM Clofazamine11 μM Yes Rifabutin 26.5 μM Brivudine 75.0 μM Clofazamine 8.8 μM YesRifabutin 21.2 μM Brivudine 60.0 μM Clofazamine 6.6 μM No Rifabutin15.91 μM Brivudine 45.0 μM Clofazamine 4.4 μM No Rifabutin 10.6 μMBrivudine 30.0 μM Clofazamine 2.2 μM No Rifabutin 5.3 μM Brivudine 15.0μM Clofazamine 1.1 μM No Rifabutin 2.65 μM Brivudine 7.5 μM Clofazamine0.66 μM No Rifabutin 1.59 μM Brivudine 4.5 μM Clofazamine 0.22 μM NoRifabutin 0.53 μM Brivudine 1.5 μM Clofazamine 0.11 μM No Rifabutin0.265 μM Brivudine 0.75 μM Rifabutin 31.8 μM No Clarithromycin 76.2 μMBrivudine 90 μM Rifabutin 26.5 μM No Clarithromycin 63.5 μM Brivudine 75μM Rifabutin 21.2 μM No Clarithromycin 50.8 μM Brivudine 60 μM Rifabutin15.9 μM No Clarithromycin 38.1 μM Brivudine 45 μM Rifabutin 10.6 μM NoClarithromycin 25.4 μM Brivudine 30 μM Rifabutin 5.3 μM NoClarithromycin 12.7 μM Brivudine 15 μM Rifabutin 2.65 μM NoClarithromycin 6.35 μM Brivudine 7.5 μM Rifabutin 1.59 μM NoClarithromycin 3.81 μM Brivudine 4.5 μM Rifabutin 0.53 μM NoClarithromycin 1.27 μM Brivudine 1.5 μM Rifabutin 0.265 μM NoClarithromycin 0.635 μM Brivudine 0.75 μM Clofazamine 13.2 μM YesClarithromycin 76.2 μM Brivudine 90 μM Clofazamine 11 μM YesClarithromycin 63.5 μM Brivudine 75 μM Clofazamine 8.8 μM YesClarithromycin 50.8 μM Brivudine 60 μM Clofazamine 6.6 μM NoClarithromycin 38.1 μM Brivudine 45 μM Clofazamine 4.4 μM NoClarithromycin 25.4 μM Brivudine 30 μM Clofazamine 2.2 μM NoClarithromycin 12.7 μM Brivudine 15 μM Clofazamine 1.1 μM NoClarithromycin 6.35 μM Brivudine 7.5 μM Clofazamine 0.66 μM NoClarithromycin 3.81 μM Brivudine 4.5 μM Clofazamine 0.22 μM NoClarithromycin 1.27 μM Brivudine 1.5 μM Clofazamine 0.11 μM NoClarithromycin 0.635 μM Brivudine 0.75 μM Rifabutin 45 μM YesClarithromycin 210 μM Rifabutin 45 μM No Clarithromycin 180 μM Rifabutin45 μM No Clarithromycin 150 μM Rifabutin 45 μM No Clarithromycin 120 μMRifabutin 45 μM No Clarithromycin 90 μM Rifabutin 45 μM NoClarithromycin 75 μM Rifabutin 60 μM Yes Clarithromycin 210 μM Rifabutin60 μM Yes Clarithromycin 180 μM Rifabutin 60 μM Yes Clarithromycin 150μM Rifabutin 60 μM Yes Clarithromycin 120 μM Rifabutin 60 μM NoClarithromycin 90 μM Rifabutin 60 μM No Clarithromycin 75 μM Rifabutin31.8 μM Yes Clarithromycin 76.2 μM ABC29460 100 μM Rifabutin 31.8 μM YesClarithromycin 76.2 μM ABC29460 80 μM Rifabutin 31.8 μM YesClarithromycin 76.2 μM ABC29460 60 μM Rifabutin 31.8 μM YesClarithromycin 76.2 μM ABC29460 40 μM Rifabutin 31.8 μM YesClarithromycin 76.2 μM ABC29460 20 μM Rifabutin 31.8 μM YesClarithromycin 76.2 μM ABC29460 10 μM Rifabutin 31.8 μM YesClarithromycin 76.2 μM ABC29460 5 μM Rifabutin 31.8 μM YesClarithromycin 76.2 μM ABC29460 2.5 μM

Based on these results, specific concentrations and combinations ofdrugs were tested to determine whether or not they were capable ofinhibiting replication of Ebola virus.

The Table below provides a summary of the concentration of drugs anddrug combinations that were found to be both non-toxic and provide areduction in the infection of Ebola virus on primary macrophage cells.

MOI = 0.01, 0.1 and 1.0 Clofazamine 6 μm Rifabutin 60 μm Clarithromycin210 μm + Brivudine 270 μm Rifabutin 31.8 μm + Clarithromycin 76.2 μmRifabutin 31.8 μm + Clarithromycin 76.2 μm + Brivudine 90 μm Rifabutin31.8 μm + Clarithromycin 76.2 μm + Brivudine 150 μm Rifabutin 31.8 μm +Clarithromycin 76.2 μm + Brivudine 210 μm MOI = 0.1 Rifabutin 45 μm +Clarithromycin 180 μm Rifabutin 45 μm + Clarithromycin 150 μm Rifabutin45 μm + Clarithromycin 120 μm Rifabutin 45 μm + Clarithromycin 90 μmRifabutin 45 μm + Clarithromycin 75 μm Rifabutin 60 μm + Clarithromycin90 μm Rifabutin 60 μm + Clarithromycin 75 μm Clarithromycin 75 μmClarithromycin 90 μm Clarithromycin 120 μm Clarithromycin 150 μmClarithromycin 180 μm ABC294640 2.5 μm ABC294640 5.0 μm ABC294640 10.0μm MOI = 1 Rifabutin 26.5 μm + Clarithromycin 63.5 μm + Brivudine 75 μm

Clarithromycin alone can inhibit EBOV infection in primary macrophages.There appears to be a synergistic effect when rifabutin andclarithromycin are administered together.

Prophetic Examples Example 2 In Vivo ScreeningExperiments—Ebola—Nonhuman Primates

Rhesus macaques will be used to determine whether administration of anagent of the present invention or a combination of agents of the presentinvention for a suitable period of time are capable of inhibiting viralreplication, decreasing viral load, or reducing at least one symptomassociated with Ebola. The experiment will consist of the same number ofnonhuman primates (NHPs) per study arm receiving various doses of agentsof the present invention alone and agents of the present inventiontogether for a suitable period of time, beginning at least one day aftera lethal intramuscular challenge with, for example, 4,000× median tissueculture infective dose (TCID₅₀) (or 2,512 plaque-forming units (pf.u.)of EBOV-K. It is contemplated that the NHPs from each study arm will besplit up into different groups that will receive the agent or agents atdifferent time points post infection, such as 30 to 75 min (Group 1), 24hours (Group 2), 48 hours (Group 3), and 72 hours (Group 4) afterinfection, and will comprise at least one daily dose. Control animalsconsisting of the same number of NHPs as the study arm will be givenphosphate-buffered saline (PBS). Kaplan-Meier survival curves for eachGroup will be created and clinical scores and fever (rectal temperature)will be measured. In addition, viral titres (EBOC viraemia) by TCID₅₀will be measured. Blood counts and serum biochemistry, including, butnot limited to, white blood cell count, lymphocyte count, lymphocytepercentage, platelet count, neutrophil count, neutrophil percentage,alanine aminotransferase, alkaline phosphatase, blood urea nitrogen,creatinine and glucose, will be measured.

Example 3 In Vivo Screening Experiments—Marburg—Nonhuman Primates

Rhesus macaques will be used to determine whether administration of anagent of the present invention or a combination of agents of the presentinvention for a suitable period of time are capable of inhibiting viralreplication, decreasing viral load, or reducing at least one symptomassociated with Marburg. The experiment will consist of the same numberof nonhuman primates (NHPs) per study arm receiving various doses ofagents of the present invention alone and agents of the presentinvention together for a suitable period of time, beginning at least oneday after a lethal dose of MARV-Angola. It is contemplated that the NHPsfrom each study arm will be split up into different groups that willreceive the agent or agents at different time points post infection,such as 30 to 75 min (Group 1), 24 hours (Group 2), 48 hours (Group 3),and 72 hours (Group 4) after infection, and will comprise at least onedaily dose. Control animals consisting of the same number of NHPs as thestudy arm will be given phosphate-buffered saline (PBS). Kaplan-Meiersurvival curves for each Group will be created and clinical scores andfever (rectal temperature) will be measured. In addition, viral titresby TCID₅₀ will be measured. Blood counts and serum biochemistry,including, but not limited to, white blood cell count, lymphocyte count,lymphocyte percentage, platelet count, neutrophil count, neutrophilpercentage, alanine aminotransferase, alkaline phosphatase, blood ureanitrogen, creatinine and glucose, will be measured.

A method of the present invention includes administering a compound toan individual infected with or exposed to a filovirus, wherein the stepof administering is carried out for a suitable time period so that theindividual is treated, and wherein the compound is represented byformula I:

-   -   or a pharmaceutically acceptable salt thereof, wherein:    -   R₁ is phenyl, 4-chlorophenyl or 4-fluorophenyl,    -   R₂ is 4-pyridyl, optionally substituted with up to 4 groups that        are independently (C₁-C₆) alkyl, halogen, haloalkyl,        —OC(O)(C₁-C₆ alkyl), —C(O)O(C₁-C₆ alkyl), —CONR′R″, —OC(O)NR′R″,        —NR′C(O)R″, —CF₃, —OCF₃, —OH, C₁-C₆ alkoxy, hydroxyalkyl, —CN,        —CO₂H, —SH, —S-alkyl, —SOR′R″, —SO₂R′, —NO₂, or NR′R″, wherein        R′ and R″ are independently H or (C₁-C₆) alkyl, and wherein each        alkyl portion of a substituent is optionally further substituted        with 1, 2, or 3 groups independently selected from halogen, CN,        OH, and NH₂,    -   R₄ is H or alkyl, and    -   n is 1 or 2; and

determining whether the individual has been treated, wherein the step ofdetermining comprising one of measuring an inhibition in viralreplication, measuring a decrease in viral load, or reducing at leastone symptom associated with the filovirus. In an embodiment, thecompound of formula I is:

In an embodiment, an effective amount of the compound of formula I isbetween about 15.0 mg/kg/day to about 20 mg/kg/day. In an embodiment,the determining step includes measuring, at at least two different timesduring the suitable time period, the viral load using a nucleic acidamplification based test. In an embodiment, the inhibition in viralreplication or the decrease in viral load is at least 10% as determinedusing a nucleic acid amplification based test. In an embodiment, theindividual is a human. In an embodiment, the filovirus is Ebola virus orMarburg virus. In an embodiment, the filovirus is Ebola virus. In anembodiment, the compound of formula I is present as a solid dosage form.In an embodiment, the solid dosage form is a capsule. In an embodiment,the method further includes administering at least one antibiotic to theindividual infected with or exposed to the filovirus for the suitabletime period, wherein the combination of the at least one antibiotic andthe compound of formula I produce a synergistic effect. In anembodiment, the at least one antibiotic is selected from one ofclarithromycin or rifabutin.

A method of the present invention includes administering at least twoantibiotics to an individual infected with or exposed to a filovirus,wherein the step of administering is carried out for a suitable timeperiod so that the individual is treated; and determining whether theindividual has been treated, wherein the step of determining includesone of measuring an inhibition in viral replication, measuring adecrease in viral load, or reducing at least one symptom associated withthe filovirus. In an embodiment, at least one of the antibiotics is amacrolide antibiotic. In an embodiment, at least one of the antibioticsis a rifamycin antibiotic. In an embodiment, the antibiotics areclarithromycin and rifabutin. In an embodiment, an effective amount ofclarithromycin is between about 12.0 mg/kg/day to about 17.0 mg/kg/day.In an embodiment, an effective amount of rifabutin is between about 4.0mg/kg/day to about 8.0 mg/kg/day. In an embodiment, the determining stepincludes measuring, at at least two different times during the suitabletime period, the viral load using a nucleic acid amplification basedtest. In an embodiment, the inhibition in viral replication or thedecrease in viral load is at least 10% as determined using a suitableassay. In an embodiment, the individual is a human. In an embodiment,the filovirus is Ebola virus or Marburg virus. In an embodiment, thefilovirus is Ebola virus.

All patents, patent applications, and published references cited hereinare hereby incorporated by reference in their entirety. Variousmodifications and variations of the described compositions and methodsof the invention will be apparent to those skilled in the art withoutdeparting from the scope and spirit of the invention. Although theinvention has been described in connection with specific embodiments, itwill be understood that the invention should not be unduly limited tosuch specific embodiments. Indeed, various modifications of thedescribed modes for carrying out the invention that are obvious to thoseskilled in the fields of molecular biology, medicine, immunology,pharmacology, virology, or related fields are intended to be within thescope of the invention.

What is claimed is:
 1. A method comprising: administering a compound toan individual infected with or exposed to a filovirus, wherein the stepof administering is carried out for a suitable time period so that theindividual is treated, and wherein the compound is represented byformula I:

or a pharmaceutically acceptable salt thereof, wherein: R₁ is phenyl,4-chlorophenyl or 4-fluorophenyl, R₂ is 4-pyridyl, optionallysubstituted with up to 4 groups that are independently (C₁-C₆) alkyl,halogen, haloalkyl, —OC(O)(C₁-C₆ alkyl), —C(O)O(C₁-C₆ alkyl), —CONR′R″,—OC(O)NR′R″, —NR′C(O)R″, —CF₃, —OCF₃, —OH, C₁-C₆ alkoxy, hydroxyalkyl,—CN, —CO₂H, —SH, —S-alkyl, —SOR′R″, —SO₂R′, —NO₂, or NR′R″, wherein R′and R″ are independently H or (C₁-C₆) alkyl, and wherein each alkylportion of a substituent is optionally further substituted with 1, 2, or3 groups independently selected from halogen, CN, OH, and NH₂, R₄ is Hor alkyl, and n is 1 or 2; and determining whether the individual hasbeen treated, wherein the step of determining comprising one ofmeasuring an inhibition in viral replication, measuring a decrease inviral load, or reducing at least one symptom associated with thefilovirus.
 2. The method of claim 1, wherein the compound of formula Iis:


3. The method of claim 1, wherein the determining step comprisesmeasuring, at at least two different times during the suitable timeperiod, the viral load using a nucleic acid amplification based test. 4.The method of claim 1, wherein the inhibition in viral replication orthe decrease in viral load is at least 10% as determined using a nucleicacid amplification based test.
 5. The method of claim 1 wherein theindividual is a human.
 6. The method of claim 1, wherein the filovirusis Ebola virus or Marburg virus.
 7. The method of claim 1, wherein thefilovirus is Ebola virus.
 8. The method of claim 1, wherein the compoundof formula I is present as a solid dosage form.
 9. The method of claim8, wherein the solid dosage form is a capsule.
 10. The method of claim1, further comprising administering at least one antibiotic to theindividual infected with or exposed to the filovirus for the suitabletime period, wherein the combination of the at least one antibiotic andthe compound of formula I produce a synergistic effect.
 11. The methodof claim 10, wherein the at least one antibiotic is selected from one ofclarithromycin or rifabutin.
 12. A method comprising: administering atleast two antibiotics to an individual infected with or exposed to afilovirus wherein the step of administering is carried out for asuitable time period so that the individual is treated; and determiningwhether the individual has been treated, wherein the step of determiningcomprising one of measuring an inhibition in viral replication,measuring a decrease in viral load, or reducing at least one symptomassociated with the filovirus.
 13. The method of claim 12, wherein atleast one of the antibiotics is a macrolide antibiotic.
 14. The methodof claim 12, wherein at least one of the antibiotics is a rifamycinantibiotic.
 15. The method of claim 12, wherein the antibiotics areclarithromycin and rifabutin.
 16. The method of claim 12, wherein thedetermining step comprises measuring, at at least two different timesduring the suitable time period, the viral load using a nucleic acidamplification based test.
 17. The method of claim 12, wherein theinhibition in viral replication or the decrease in viral load is atleast 10% as determined using a suitable assay.
 18. The method of claim12, wherein the individual is a human.
 19. The method of claim 12,wherein the filovirus is Ebola virus or Marburg virus.
 20. The method ofclaim 12, wherein the filovirus is Ebola virus.